TITLE: Gene Expression in Algae and Fungi Including Yeast
PUBLICATION DATE:January, 1993
ENTRY DATE: March, 1994
EXPIRATION DATE: None
UPDATE: As needed
CONTACT: Biotechnology Information Center(biotech@nalusda.gov)
National Agricultural Library
DOCUMENT TYPE: Text
DOCUMENT SIZE: 232.9k, approx. 129 pp.
ISSN: 1052-5378
United States Department of Agriculture
National Agricultural Library
10301 Baltimore Blvd.
Beltsville, Maryland 20705-2351
Gene Expression in Algae and Fungi Including Yeast January 1991 - November 1992
QB 93-10
137 citations from Agricola
Janet Saunders and Robert Warmbrodt
Biotechnology Information Center
January 1993 National Agricultural Library Cataloging Record:
Saunders, Janet
Gene expression in algae and fungi, including yeast.
(Quick bibliography series ; 93-10)
SEARCH STRATEGY
Set Items Description
S1 5992 EXPRESS?/TI
S2 99924 ALGA? OR DEUTEROMYCOTINA OR FUNG? OR MOLD? OR
MOULD? OR YEAST?
S3 705 S1 AND S3
S4 166 S3 AND PY=1991:1992
Gene Expression in Algae and Fungi, Including Yeast
1 NAL Call. No.: 500 N21P
ACE1, a copper-dependent transcription factor, activates
expression of the yeast copper, zinc superoxide dismutase
gene.
Gralla, E.B.; Thiele, D.J.; Silar, P.; Valentine, J.S.
Washington, D.C. : The Academy; 1991 Oct01.
Proceedings of the National Academy of Sciences of the United
States of America v. 88 (19): p. 8558-8562; 1991 Oct01.
Includes references.
Language: English
Descriptors: Saccharomyces cerevisiae; Yeasts; Binding site; Copper; Enzyme activity; Gene expression; Metal ions; Superoxide dismutase; Transcription; Zinc
Abstract: Copper, zinc superoxide dismutase (SOD1 gene product) (superoxide:superoxide oxidoreductase, EC 1.15.1.1) is a copper-containing enzyme that functions to prevent oxygen toxicity. In the yeast Saccharomyces cerevisiae, copper levels exert some control over the level of SOD1 expression. We show that the ACE1 transcriptional activator protein, which is responsible for the induction of yeast metallothionein (CUP1) in response to copper, also controls the SOD1 response to copper. A single binding site for ACE1 is present in the SOD1 promoter region, as demonstrated by DNase I protection and methylation interference experiments, and is highly homologous to a high-affinity ACE1 binding site in the CUP1 promoter. The functional importance of this DNA-protein interaction is demonstrated by the facts that (i) copper induction of SOD1 mRNA does not occur in a strain lacking ACE1 and (ii) it does not occur in a strain containing a genetically engineered SOD1 promoter that lacks a functional ACE1 binding site.
2 NAL Call. No.: QH573.N37
The acid phosphatase of Saccharomyces cerevisiae: a model to
study wall protein expression.
Monod, M.; Haguenauer-Tsapis, R.; Silve, S.; Togni, G.;
Hinnen, A. Berlin, W. Ger. : Springer-Verlag; 1991.
NATO ASI series : Series H : Cell biology v. 53: p. 257-268;
1991. In the series analytic: Fungal cell wall and immune
response / edited by J.P. Latge and D. Boucias. Proceedings of
the NATO Advanced Research Workshop, September 29-October 5,
1990, Elounda, Greece. Includes references.
Language: English
Descriptors: Saccharomyces cerevisiae; Acid phosphatase; Enzyme activity; Protein synthesis; Cell wall components; Plant proteins; Endoplasmic reticulum
3 NAL Call. No.: QH426.C8
Altered expression of the steroid bioconverting pathway in pAN
7-1 transformants of Cochliobolus lunatus.
Rozman, D.; Komel, R.
Berlin, W. Ger. : Springer International; 1991.
Current genetics v. 20 (5): p. 385-389; 1991. Includes
references.
Language: English
Descriptors: Cochliobolus lunatus; Escherichia coli; Aspergillus nidulans; Genetic transformation; Plasmids; Vectors; Gene expression; Progesterone; Steroid metabolism; Androstenedione; Testosterone
Abstract: The filamentous fungus C. lunatus converts progesterone mainly to its 11 beta-hydroxy derivative. C. lunatus transformed with the plasmid pAN 7-1, which contains the E. coli hph gene expressed under the control of the A. nidulans gpd and trpC expression signals, lacks this activity, but exhibits acetyl side chain degradation of progesterone through the reaction scheme progesterone leads to 20 betahydroxy -progesterone leads to delta 4-androstene-3,17-dione leads to testolactone + testosterone. The main part of this metabolic pathway is not expressed in the non-transformed strain. It was determined that the site-specific integration of the plasmid into the genome directly influences the expression of genes involved in the bioconversion of steroids.
4 NAL Call. No.: QH442.A1G4 An androgen-inducible expression system for Saccharomyces cerevisiae. Purvis, I.J.; Chotai, D.; Dykes, C.W.; Lubahn, D.B.; French, F.S.; Wilson, E.M.; Hobden, A.N. Amsterdam : Elsevier Science Publishers; 1991. Gene v. 106 (1): p. 35-42; 1991. Includes references.
Language: English
Descriptors: Saccharomyces cerevisiae; Vectors; Gene expression; Androgens; Receptors; Genes; Man; Promoters; Recombinant DNA
Abstract: A novel controllable expression system for Saccharomyces cerevisiae has been developed. Expression of the gene encoding the human androgen receptor, from a strong yeast promoter, results in transactivation of a hybrid promoter carrying androgen-responsive sequences such that a target gene may be expressed in an androgen-dependent manner. By selection of an appropriate combination of androgen receptor level, target-gene copy number and concentration of the androgenic ligand, dihydrotestosterone, the expression level can be set within a 1400-fold range with no detectable effect on normal cell growth.
5 NAL Call. No.: 442.8 Z34 Association of cytochrome b translational activator proteins with the mitochondrial membrane: implications for cytochrome b expression in yeast. Michaelis, U.; Korte, A.; Rodel, G. Berlin, W. Ger. : Springer International; 1991 Nov. M G G : Molecular and general genetics v. 230 (1/2): p. 177-185; 1991 Nov. Includes references.
Language: English
Descriptors: Saccharomyces cerevisiae; Translation; Gene expression; Cytochrome b; Messenger RNA; Regulation; Genes; Plant proteins; Mitochondria; Plasma membranes
Abstract: The products of the nuclear genes CBS1 and CBS2 are both required for translational activation of mitochondrial apocytochrome b in yeast. We report the intramitochondrial localization of both proteins by use of specific antisera. Based on its solubilization properties the CBS1 protein is presumed to be a component of the mitochondrial membrane; the detergent concentrations needed to release CBS1 from mitochondria are almost the same as for cytochrome c1. In contrast, CBS2 behaves like a soluble protein, with some characteristics of a membrane-associated protein. A model is presented for translational activation of cytochrome b, which might also be applicable to translational regulation of other mitochondrial genes.
6 NAL Call. No.: 381 J824
The Candida albicans myristoyl-CoA:protein Nmyristoyltransferase
gene. Isolation and expression in
Saccharomyces cerevisiae and Escherichia coli. Wiegand, R.C.;
Carr, C.; Minnerly, J.C.; Pauley, A.M.; Carron, C.P.; Langner,
C.A.; Duronio, R.J.; Gordon, J.I.
Baltimore, Md. : American Society for Biochemistry and
Molecular Biology; 1992 Apr25.
The Journal of biological chemistry v. 267 (12): p. 8591-8598;
1992 Apr25. Includes references.
Language: English
Descriptors: Candida albicans; Myristic acid; Coenzyme a; Transferases; Structural genes; Isolation; Gene expression; Nucleotide sequences; Amino acid sequences
Abstract: Myristoyl-CoA:protein N-myristoyltransferase (NMT) has recently been identified as a target for antiviral and antifungal therapy. Candida albicans is a dimorphic, asexual yeast that is a major cause of systemic fungal infections in immunosuppressed humans. metabolic labeling studies indicate that C. albicans synthesizes one principal 20-kDa N-myristoylprotein. The single copy C. albicans NMT gene (ca-NMT1) was isolated and encodes a 451-amino acid protein that has 55% identity with Saccharomyces cerevisiae NMT. C. albicans NMT1 is able to complement the lethal phenotype of S. cerevisiae nmt1 null mutants by directing efficient acylation of the approximately 12 endogenous N-myristoylproteins produced by S. cerevisiae. C. albicans NMT was produced in Escherichia coli, a prokaryote with no endogenous NMT activity. In vitro studies of purified E. coli-derived S. cerevisiae and C. albicans NMTs revealed species-specific differences in the kinetic properties of synthetic octapeptide substrates derived from known N-myristoylproteins. Together these data indicate that C. albicans and S. cerevisiae NMTs have similar yet distinct substrate specificities which may be of therapeutic significance.
7 NAL Call. No.: 442.8 Z34
The CCR1 (SNF1) and SCH9 protein kinases act independently of
cAMP-dependent protein kinase and the transcriptional
activator ADR1 in controlling yeast ADH2 expression.
Denis, C.L.; Audino, D.C.
Berlin, W. Ger. : Springer International; 1991 Oct.
M G G : Molecular and general genetics v. 229 (3): p. 395-399;
1991 Oct. Includes references.
Language: English
Descriptors: Saccharomyces cerevisiae; Gene expression; Alcohol dehydrogenase; Genes; Transcription; Genetic regulation; Protein kinase; C-amp; Dna binding proteins; Enzyme activity
Abstract: cAMP-dependent protein kinase (cAPK) is implicated in the inactivation of the yeast transcriptional activator ADR1, which regulates glucose-repressible ADH2 gene expression. The interdependence of cAPK, SCH9 (a protein kinase that when overexpressed can functionally substitute for cAPK), and the CCR1 (SNF1) protein kinase that is required for ADH2 expression was studied. SCH9 was found to be required for ADH2 expression in contrast to the inhibitory role played by cAPK. CCR1 and SCH9 were observed to affect ADH2 expression independently of both ADR1 and cAPK. In contrast, cAPK was shown to exert its effects on ADH2 solely through ADR1. These results indicate that the SCH9 and CCR1 protein kinases are components of regulatory pathways separate from that utilized by cAPK to control ADR1 activity and ADH2 expression.
8 NAL Call. No.: QH426.C8
Characterization of a novel open reading frame, urf a, in the
mitochondrial genome of fission yeast: Correlation of urf a
mutations with a mitochondrial mutator phenotype and a
possible role of frameshifting in urf a expression. Zimmer,
M.; Krabusch, M.; Wolf, K.
Berlin, W. Ger. : Springer International; 1991.
Current genetics v. 19 (2): p. 95-102; 1991. Includes
references.
Language: English
Descriptors: Endomycetales; Mitochondrial DNA; Mitochondrial genetics; Genes; Mutations; Genomes; Nucleotide sequences; Phenotypes; Mutator genes; Segregation; Translation; Cytoplasmic inheritance; Amino acid sequences
Abstract: Between the genes for tRNA(gln) and tRNA(ile) an open reading frame of 227 amino acids has been identified which is unique among known mitochondrial genomes and which has been termed urf a (Lang et al. 1983; Kornrumpf et al. 1984). It uses the "mitochondrial" genetic code, i.e., it contains a TGA codon, whereas all other protein-encoding genes, and all but one intronic open reading frame, use the "standard" genetic code (UGG for tryptophan). A previous paper has demonstrated that "mutator" strains show an increased formation of mitochondrial drug-resistant and respirationdeficient mutants (including deletions). In this paper we show that the mutator activity is correlated with mutations in urf
9 NAL Call. No.: QP601.M49
Characterization of human P450IIC isozymes by using yeast
expression system. Kato, R.; Yasumori, T.; Yamazoe, Y.
New York, N.Y. : Academic Press; 1991.
Methods in enzymology v. 206: p. 183-190; 1991. In the series
analytic: Cytochrome P450 / edited by M.R. Waterman and E.F.
Johnson. Includes references.
Language: English
Descriptors: Cytochrome p-450; Isoenzymes; Vectors; Gene expression; Genetic transformation; Man; Saccharomyces cerevisiae
10 NAL Call. No.: QH442.A1G4 Characterization of INF56, a gene expressed during infection structure development of Uromyces appendiculatus. Xuei, X.; Bhairi, S.; Staples, R.C.; Yoder, O.C. Amsterdam : Elsevier Science Publishers; 1992. Gene v. 110 (1): p. 49-55; 1992. Includes references.
Language: English
Descriptors: Uromyces appendiculatus; Genes; Introns; Polypeptides; Nucleotide sequences; Amino acid sequences; Gene expression; Messenger RNA; Hyphae; Cell differentiation; Infections; Pathogenesis; Thigmotropism
Abstract: The bean rust fungus, Uromyces appetidiculatus, undergoes thigmotropic differentiation to produce infection structures. Six differentiation-specific genes have been isolated and one, INF24, has been characterized [Bhairi et al., Gene 81 (1989) 237-243]. Here, we report the structure of a second gene, IVF56, which was subcloned on a 2.6-kb fragment and sequenced. The location of the 1.0-kb INF56 transcript was determined by S1 nuclease protection and primer extension. A TATA box was found 38 bp upstream and a CAAT box 130 bp upstream from the major transcription start point (tsp). The gene contains two open reading frames: ORF2 is nested within ORF1; they share a 67-bp intron. ORF1 encodes a 14.1-kDa polypeptide which has an amino acid sequence rich in Gly, Pro and Ser. It has sequence similarity to a functional domain (V2) of mammalian cytokeratin type II. ORF2 encodes a 10.1-kDa polypeptide which is rich in Pro. It shares similarity with the cell-surface recognition region of chicken fibronectin. Hybrid selection and in vitro translation of the INF56 mRNA yielded two polypeptides of 15.5 and 23 kDa, as estimated by sodium dodecyl sulfate-polyacrylamide-gel electrophoresis. INF56 is constitutively expressed at a low level, but the abundance of its steady-state transcript is upshifted 4.5 h after spore hydration during the period that infection structures are formed.
11 NAL Call. No.: QP501.B642
Chromatin structure in a region of a yeast transposable
element regulating adjacent gene expression.
Feaver, W.J.; Pearlman, R.E.
Ottawa : National Research Council of Canada; 1991 May.
Biochemistry and cell biology; Biochimie et biologie
cellulaire v. 69 (5/6): p. 392-398; 1991 May. Includes
references.
Language: English
Descriptors: Saccharomyces cerevisiae; Mutants; Nuclei; Chromatin; Genes; Transposable elements; Gene expression; Genetic regulation; Dna sequencing
12 NAL Call. No.: 448.3 AP5
Chromosomal integration and expression of two bacterial alphaacetolactate
decarboxylase genes in brewer's yeast.
Blomqvist, K.; Suihko, M.L.; Knowles, J.; Penttila, M.
Washington, D.C. : American Society for Microbiology; 1991
Oct. Applied and environmental microbiology v. 57 (10): p.
2796-2803; 1991 Oct. Includes references.
Language: English
Descriptors: Saccharomyces cerevisiae; Strains; Carboxylyases; Genes; Gene expression; Genetic transformation; Klebsiella; Enterobacter aerogenes; Diacetyl; Metabolism; Enzyme activity; Beers; Brewing industry
Abstract: A bacterial gene encoding alpha-acetolactate decarboxylase, isolated from Klebsiella terrigena or Enterobacter aerogenes, was expressed in brewer's yeast. The genes were expressed under either the yeast phosphoglycerokinase (PGKI) or the alcohol dehydrogenase (ADHI) promoter and were integrated by gene replacement by using cotransformation into the PGKI or ADHI locus, respectively, of a brewer's yeast. The expression level of the alpha-acetolactate decarboxylase gene of the PGK1 integrant strains was higher than that of the ADH1 integrants. Under pilot-scale brewing conditions, the alpha-acetolactate decarboxylase activity of the PGK1 integrant strains was sufficient to reduce the formation of diacetyl below the taste threshold value, and no lagering was needed. The brewing properties or the recombinant yeast strains were otherwise unaltered, and the quality (most importantly, the flavor) of the trial beers produced was as good as that of the control beer.
13 NAL Call. No.: 381 J824
Cloning and expression in Escherichia coli of the gene
encoding Aspergillus flavus urate oxidase.
Legoux, R.; Delpech, B.; Dumont, X.; Guillemot, J.C.; Ramond,
P.; Shire, D.; Caput, D.; Ferrara, P.; Loison, G.
Baltimore, Md. : American Society for Biochemistry and
Molecular Biology; 1992 Apr25.
The Journal of biological chemistry v. 267 (12): p. 8565-8570;
1992 Apr25. Includes references.
Language: English
Descriptors: Aspergillus flavus; Uric acid; Oxidoreductases; Structural genes; Cloning; Gene expression; Nucleotide sequences; Amino acid sequences
Abstract: Amino acid sequencing of peptides obtained after proteolytic hydrolysis of Aspergillus flavus urate oxidase (uricase) permitted the design of oligodeoxynucleotide probes that were used to obtain 1.2- and 5-kilobase pair DNA fragments from A. flavus cDNA and genomic libraries, respectively. The cDNA fragment contained the entire coding region for uricase, and comparison with the genomic fragment revealed the presence of two short introns in the coding region of the gene. A. flavus uricase has around 40% overall identity with uricases from higher organisms but with many conserved amino acids. Hitherto highly conserved consensus patterns found in other uricases were found to be modified in the A. flavus enzyme, notably the sequence Val-Leu-Lys-ThrThr -Gln-Ser near position 150, which in the filamentous fungus is uniquely modified to Val-Leu-Lys-Ser-Thr-Asn-Ser. Silent mutations were introduced by cassette mutagenesis near the 5'- extremity of the coding sequence in order to conform with Escherichia coli codon usage, and the uricase was expressed in the E. coli cytoplasm in a completely soluble, biologically active form.
14 NAL Call. No.: QH442.A1G4 Cloning and expression in Saccharomyces cerevisiae of the NAD(P)H-dependent xylose reductase-encoding gene (XYL1) from the xylose-assimilating yeast Pichia stipitis. Amore, R.; Kotter, P.; Kuster, C.; Ciriacy, M.; Hollenberg, C.P. Amsterdam : Elsevier Science Publishers; 1991. Gene v. 109 (1): p. 89-97; 1991. Includes references.
Language: English
Descriptors: Saccharomyces cerevisiae; Pichia stipitis; Genes; Cloning; Oxidoreductases; Xylose; Nad; Nadh; Nucleotide sequences; Amino acid sequences; Genetic transformation; Gene expression; Enzyme activity
Abstract: The XYL1 gene of the yeast Pichia stipitis has been isolated from a genomic library using a specific cDNA probe, and its nucleotide (nt) sequence has been determined. In the 5' noncoding region of the P. stipitis XYL1 gene a TATAAA element (known to be necessary for transcription initiation in most yeast genes) is found at nt -81, and two CCAAT recognition motifs (often referred to as the CCAAT box) are present at nt -146 and -106. The XYL1 encodes a polypeptide of 35927 Da that constitutes a NADH/NADPH-dependent xylose reductase (XR). The enzyme is part of the xylose-xylulose pathway that is absent or only weakly expressed in Saccharomyces cerevisiae. Extensive homology is found to the N terminus of the XR of Pachysolen tannophilus and Candida shehatae. None of the known cofactor binding domains found in many NAD-dependent dehydrogenases are present in the protein. Transformants of S. cerevisiae containing XYL1 of P. stipitis synthesize an active XR. Fusion of XYL1 with the prokaryotic tac promoter leads to a gene that can be expressed in S. cerevisiae and Escherichia coli.
15 NAL Call. No.: 442.8 Z34
Cloning and expression in yeast of a cDNA clone encoding
Aspergillus oryzae neutral protease II, a unique
metalloprotease.
Tatsumi, H.; Murakami, S.; Tsuji, R.F.; Ishida, Y.; Murakami,
K.; Masaki, A.; Kawabe, H.; Arimura, H.; Nakano, E.; Motai, H.
Berlin, W. Ger. : Springer International; 1991 Aug.
M G G : Molecular and general genetics v. 228 (1/2): p.
97-103; 1991 Aug. Includes references.
Language: English
Descriptors: Aspergillus oryzae; Saccharomyces cerevisiae; Genes; Proteinases; Cloning; Nucleotide sequences; Amino acid sequences; Genetic transformation; Transgenics; Gene expression; Enzyme precursors; Zinc; Binding site; Enzyme activity
Abstract: The neutral protease II (NpII) from Aspergillus oryzae is a zinc-containing metalloprotease with some unique properties. To elucidate its structure, we isolated a fulllength cDNA clone for NpII. Sequence analysis reveals that NpII has a prepro region consisting of 175 amino acids preceding the mature region, which consists of 177 amino acids. As compared with other microbial metalloproteases, NpII is found to be unique in that it shares only a limited homology with them around two zinc ligand His residues and that the positions of the other zinc ligand (Glu) and the active site (His) cannot be established by homology. When a plasmid designed to express the prepro NpII cDNA was introduced into Saccharomyces cerevisiae and the transformant was cultured in YPD medium (2% glucose, 2% polypeptone, 1% yeast extract), it secreted a proNpII. However, in a culture of the same medium containing 0.2 mM ZnCl2, it secreted a mature NpII with a specific activity and N-terminus identical to those of native NpII. This observation suggests that either an autoproteolytic activity or a yeast protease effected the processing.
16 NAL Call. No.: 470 SCI2
Cloning and expression in yeast of a plant potassium ion
transport system. Sentenac, H.; Bonneaud, N.; Minet, M.;
Lacroute, F.; Salmon, J.M.; Gaymard, F.; Grignon, C.
Washington, D.C. : American Association for the Advancement of
Science; 1992 May01.
Science v. 256 (5057): p. 663-665; 1992 May01. Includes
references.
Language: English
Descriptors: Arabidopsis thaliana; Gene expression; Cloning; Saccharomyces cerevisiae; Potassium; Nucleotide sequences; Amino acid sequences
Abstract: A membrane polypeptide involved in K+ transport in a higher plant was cloned by complementation of a yeast mutant defective in K+ uptake with a complementary DNA library from Arabidopsis thaliana. A 2.65-kilobase complementary DNA conferred ability to grow on media with K+ concentration in the micromolar range and to absorb K+ (or (86)Rb+) at rates similar to those in wild-type yeast. The predicted amino acid sequence (838 amino acids) has three domains: a channelforming region homologous to animal K+ channels, a cyclic nucleotide-binding site, and an ankyrin-like region.
17 NAL Call. No.: QH426.C8 Cloning and expression of a chitinase gene from the hyperparasitic fungus Aphanocladium album. Blaiseau, P.L.; Kunz, C.; Grison, R.; Bertheau, Y.; Brygoo, Y. Berlin, W. Ger. : Springer International; 1992. Current genetics v. 21 (1): p. 61-66; 1992. Includes references.
Language: English
Descriptors: Deuteromycotina; Fungal antagonists; Fusarium oxysporum; Hyperparasitism; Genes; Chitinase; Recombinant DNA; Gene expression; Messenger RNA; Genetic transformation; Nucleotide sequences; Restriction mapping; Genetic regulation; Amino acid sequences
Abstract: Recombinant clones from a cDNA library of an Aphanocladium album chitinase-overproducing mutant strain were isolated by screening with antiserum against a 39 kDa chitinase purified from this hyperparasitic fungus. Analysis of the isolated positive clones indicated that most of them carried the same cDNA. A cDNA from this group was used as a hybridization probe to isolate an 8 kb DNA fragment from a genomic library of the wild-type strain. The chitinase 1 gene was mapped to this fragment by two independent approaches. Its partial DNA sequence was in perfect agreement with an aminoterminal peptide sequence obtained by sequencing 23 amino acids of the 39 kDa chitinase. Its transfer in Fusarium oxysporum resulted in a transformant producing both a protein of about 39 kDa that cross-reacted with the chitinase antiserum and a chitinase activity that was inhibited by the same antiserum. Northern blot analysis indicates that the cloned chitinase gene was subject to catabolite repression and appeared inducible by chitin.
18 NAL Call. No.: 381 J824
The cloning and expression of a gene encoding old yellow
enzyme from Saccharomyces carlsbergensis.
Saito, K.; Thiele, D.J.; Davio, M.; Lockridge, O.; Massey, V.
Baltimore, Md. : American Society for Biochemistry and
Molecular Biology; 1991 Nov05.
The Journal of biological chemistry v. 266 (31): p.
2070-20724; 1991 Nov05. Includes references.
Language: English
Descriptors: Saccharomyces uvarum; Nadph; Oxidoreductases; Gene expression; Cloning; Recombinant DNA; Nucleotide sequences; Amino acid sequences
Abstract: We have identified a gene that encodes Old Yellow Enzyme in brewer's bottom yeast. The open reading frame encodes a polypeptide of 400 amino acids with Mr = 45,021. Using the T7 RNA polymerase system, recombinant enzyme was expressed in Escherichia coli. 17 mg of Old Yellow Enzyme was obtained from a 3-liter cell culture, and the recombinant enzyme had NADPH oxidase activity. On fast protein liquid chromatography separation, the recombinant enzyme showed a single large peak, while native enzyme from brewer's bottom yeast separated into five fractions on fast protein liquid chromatography. Southern blot analysis showed that there are at least two Old Yellow Enzyme genes in brewer's bottom yeast genomic DNA. These results suggest that the heterogeneity of Old Yellow Enzyme in Saccharomyces carlsbergensis is due to the presence of multiple genes.
19 NAL Call. No.: 381 J824
Cloning and expression of a yeast protein tyrosine
phosphatase. Guan, K.; Deschenes, R.J.; Qiu, H.; Dixon, J.E.
Baltimore, Md. : American Society for Biochemistry and
Molecular Biology; 1991 Jul15.
The Journal of biological chemistry v. 266 (20): p.
12964-12970; 1991 Jul15. Includes references.
Language: English
Descriptors: Saccharomyces cerevisiae; Tyrosine; Phosphoric monoester hydrolases; Phosphorylation; Polymerase chain reaction; Cloning; Nucleotide sequences; Amino acid sequences
Abstract: To study the regulation of tyrosine phosphorylation/dephosphorylation in Saccharomyces cerevisiae, a protein tyrosine phosphatase (PTPase) was cloned by the polymerase chain reaction (PCR). Conserved amino acid sequences within the mammalian PTPases were used to design primers which generated a yeast PCR fragment. The sequence of the PCR fragment encoded a protein with homology to the mammalian PTPases. The PCR fragment was used to identify the yeast PTP1 gene which has an open reading frame encoding a 335-amino acid residue protein. This yeast PTPase shows 26% sequence identity to the rat PTPase, although highly conserved residues within the mammalian enzymes are invariant in the yeast protein. The yeast PTP1 is physically linked to the 5'- end of a heat shock gene SSB1. This yeast PTP1 gene was expressed in Escherichia coli and obtained in a highly purified form by a single affinity chromatography step. The recombinant yeast PTPase hydrolyzed phosphotyrosine containing substrates approximately 1000 times faster than a phosphoserine containing substrate. Gene disruption of yeast PTP1 has no visible effect on vegetative growth.
20 NAL Call. No.: QP501.E8
Cloning and sequencing of arg3 and agr11 genes of
Schizosaccharomyces pombe on a 10-kb DNA fragment.
Heterologous expression and mitochondrial targeting of their
translation products.
Huffel, C. van; Dubois, E.; Messenguy, F.
New York, NY : Springer-Verlag New York Inc; 1992 Apr.
European journal of biochemistry v. 205 (1): p. 33-43; 1992
Apr. Includes references.
Language: English
Descriptors: Yeasts; Saccharomyces cerevisiae; Mutants; Strains; Arginine; Anabolism; Genes; Genetic code; Gene expression; Clones; Promoters; Nucleotide sequences; Amino acid sequences; Acetylglutamic acid; Kinases; Oxidoreductases; Ornithine carbamoyltransferase; Enzyme activity; Translation; Mitochondria
Abstract: The Schizosaccharomyces pombe arginine anabolic
genes encoding ornithine carbamoyltransferase (arg3) and
acetylglutamate kinase/acetylglutamyl-phosphate reductase
(arg11) were cloned by functional complementation of S. pombe
arg3 and arg11 mutant strains from S. pombe DNA genomic
libraries. Restriction analysis and sequencing of the two
clones showed that both genes are located on a common DNA
fragment. The arg3 gene encodes a 327-amino-acid polypeptide
presenting a strong identity to Saccharomyces cerevisiae and
human ornithine carbamoyltransferases. The arg11 gene encodes
a 884-amino-acid polypeptide. The acetylglutamate kinase and
acetylglutamate-phosphate reductase domains have been defined
by their identity with the S. cerevisiae ARG5,6 protein. The
cloned arg11 gene from S. pombe does not complement an arg5,6
mutation in S. cerevisiae, nor does the ARG5,6 gene complement
the S. pombe arg11- mutation. In contrast, both ornithinecarbamoyltransferase
-encoding genes function in S. pombe.
However, the S. pombe arg3 gene complements only weakly an
arg3 S. cerevisiae strain, which is in agreement with the low
level of expression of the S. pombe gene in S. cerevisiae. The
subcellular localization of both ornithine
carbamoyltransferases in the two yeasts indicates that, in
contrast to the S. pombe enzyme, more than 95% of the S.
cerevisiae enzyme remains in the S. pombe cytoplasm. The low
expression of S. pombe ornithine carbamoyltransferases in S.
cerevisiae did not allow its localization. The promoters of S.
pombe arg3 and arg11 genes do not present striking
similarities among themselves nor with the promoters of the
equivalent genes of S. cerevisiae.
21 NAL Call. No.: QH442.A1G4
Cloning, expression and characterization of a cDNA encoding a
lipase from Rhizopus delemar.
Haas, M.J.; Allen, J.; Berka, T.R.
Amsterdam : Elsevier Science Publishers; 1991.
Gene v. 109 (1): p. 107-113; 1991. Includes references.
Language: English
Descriptors: Rhizopus; Genes; Cloning; Triacylglycerol lipase; Nucleotide sequences; Amino acid sequences; Enzyme activity; Gene transfer; Gene expression; Escherichia coli
Abstract: A lambda-gt11 cDNA library was constructed in Escherichia coli using poly(A)-selected mRNA from the fungus, Rhizopus (Rp.) delemar. Lipase-producing members of the library were identified by means of a phenotypic score wherein the release of fatty acids by lipase causes a characteristic color change in the growth medium. One such isolate contained a 1287-bp insert (LIP cDNA) which hybridizes to 1.25- to 1.35- kb mRNA species from Rp. delemar. The lipase produced in E. coli containing the LIP cDNA exhibits the same substrate selectivity as the authentic fungal enzyme, hydrolyzing ester bonds at the stereospecific numbering (sn) sn-1 and sn-3, but not the sn-2, positions of triglycerides. The complete nucleotide sequence of the LIP cDNA was determined. By reference to the N-terminal sequence of authentic Rp. delemar lipase, the lipase-encoding region was identified within this fragment. The LIP cDNA encodes a putative preprolipase consisting of a 26-amino-acid(aa) signal sequence, a 97-aa propeptide, and a 269-aa mature enzyme. The predicted mature lipase has the same molecular weight and aa composition as that of Rp. delemar, is highly homologous to that produced by the fungus Rhizomucor miehei, and contains the consensus pentapeptide (Gly-Xaa-Ser-Yaa-Gly) which is conserved among lipolytic enzymes. It is concluded that the LIP cDNA is an essentially full-length analogue of the lipase-encoding gene of Rp. delemar. The lipase encoded by the LIP cDNA occupies a cytoplasmic location when synthesized in E. coli. Unprocessed forms of the lipase accumulate in E. coli.
22 NAL Call. No.: QH442.A1G4 Cloning, sequence analysis and transcriptional expression of a ras gene of the edible basidiomycete Lentinus edodes. Hori, K.; Kajiwara, S.; Saito, T.; Miyazawa, H.; Katayose, Y.; Shishido, K. Amsterdam : Elsevier Science Publishers; 1991. Gene v. 105 (1): p. 91-96; 1991. Includes references.
Language: English
Descriptors: Lentinula edodes; Oncogenes; Cloning; Nucleotide sequences; Transcription; Gene expression; Introns; Amino acid sequences; Developmental stages; Biological development; Camp; Mycelium
Abstract: In the edible basidiomycete, Lentinus edodes, the presence of a high level of intracellular cyclic AMP (cAMP) is closely related to the onset of fruiting and/or primordium formation. Since a close relationship between intracellular cAMP levels and expression of ras genes was reported for organisms such as Saccharomyces cerevisiae and Dictyostelium discoideum, we have cloned and sequenced a ras gene homologue from L. edodes (Le.), and analyzed its expression during development of the fungus. This gene, named Le.ras, has a coding capacity of 217 amino acids (aa) interrupted by six small introns. The deduced Le.Ras protein exhibited the highest homology to the Schizosaccharomyces pombe RAS protein (219 aa): 86% homology in the N-terminal 80-aa sequence and 74% homology in the next 80 aa. The Le.ras gene was transcribed at similar levels during mycelial development in fruiting-body formation, suggesting no direct correlation of Le.ras expression with intracellular cAMP levels in this organism.
23 NAL Call. No.: QH426.C8 Cloning, sequencing and expression of the Schwanniomyces occidentalis NADP-dependent glutamate dehydrogenase gene. De Zoysa, P.A.; Connerton, I.F.; Watson, D.C.; Johnston, J.R. Berlin, W. Ger. : Springer International; 1991. Current genetics v. 20 (3): p. 219-224; 1991. Includes references.
Language: English
Descriptors: Yeasts; Glutamate dehydrogenase; Nadp; Genes; Nucleotide sequences; Amino acid sequences; Gene expression; Saccharomyces cerevisiae
Abstract: The cloned NADP-specific glutamate dehydrogenase (GDH) genes of Aspergillus nidulans (gdhA) and Neurospora crassa (am) have been shown to hybridize under reduced stringency conditions to genomic sequences of the yeast Schwanniomyces occidentalis. Using 5' and 3' gene-specific probes, a unique 5.1 kb BclI restriction fragment that encompasses the entire Schwanniomyces sequence has been identified. A recombinant clone bearing the unique BclI fragment has been isolated from a pool of enriched clones in the yeast/E. coli shuttle vector pWH5 by colony hybridization. The identity of the plasmid clone was confirmed by functional complementation of the Saccharomyces cerevisiae gdh-1 mutation. The nucleotide sequence of the Schw. occidentalis GDH gene, which consists of 1380 nucleotides in a continuous reading frame of 459 amino acids, has been determined. The predicted amino acid sequence shows considerable homology with GDH proteins from other fungi and significant homology with all other available GDH sequences.
24 NAL Call. No.: 381 J824
Cloning, structural analysis, and expression of the glycogen
phosphorylase-2 gene in Dictyostelium.
Rutherford, C.L.; Peery, R.B.; Sucic, J.F.; Yin, Y.; Rogers,
P.V.; Luo, S.; Selmin, O.
Baltimore, Md. : American Society for Biochemistry and
Molecular Biology; 1992 Feb05.
The Journal of biological chemistry v. 267 (4): p. 2294-2302;
1992 Feb05. Includes references.
Language: English
Descriptors: Dictyostelium; Cell differentiation; Glycogen phosphorylase; Structural genes; Gene expression; Cloning; Enzyme activity; Purification; C-amp; Regulation; Nucleotide sequences; Amino acid sequences
Abstract: The glycogen phosphorylase-2 (GP2) activity that appears during the cell differentiation of Dictyostelium was purified to homogeneity. The molecular weight of the nondenatured enzyme was 200,000 as determined by Sephacryl S-300 gel filtration and was 107,000 on sodium dodecyl sulfate-polyacrylamide gel electrophoresis, suggesting that the native enzyme consists of two similar subunits. The intact protein was digested with trypsin and protease V8, and the resulting peptides were purified by microbore high pressure liquid chromatography. The peptides were sequenced, and oligonucleotides were constructed for polymerase chain reaction amplification of the GP2 gene from Dictyostelium genomic DNA template. The resulting polymerase chain reaction products were sequenced directly and were confirmed to encode portions of the GP2 gene. These fragments were used to probe a partial EcoRI genomic library for the remainder of the GP2 gene. The nucleotide sequence of the GP2-selected clones revealed an open reading frame of 2975 base pairs that was interrupted by two introns of 109 and 105 base pairs, respectively. The open reading frame encoded a protein of 992 amino acids with a calculated molecular mass of 112,500 Da and an isoelectric point of 6.4. An unusual sequence within the second exon of GP2, in which the triplet CAA was repeated 11 times, resulted in 11 in-frame glutamine residues of a possible 15 amino acids coded for by this region. The CAA repeat was transcribed, as shown by the sequence of cDNA. Comparison of the amino acid sequence of Dictyostelium GP2 to the phosphorylases from other organisms revealed that the Dictyostelium protein was 50 and 44% identical to yeast and rabbit muscle phosphorylases, respectively. Northern blot analysis showed that GP2 mRNA was absent in amebas and the early stages of development, reached a maximum level of expression at the slug stage, and then decreased in the terminal stages of development. Comparison of the mRNA expression wi
25 NAL Call. No.: 442.8 G28
The clr1 locus regulates the expression of the cryptic matingtype
loci of fission yeast.
Thon, G.; Klar, A.J.S.
Baltimore, Md. : Genetics Society of America; 1992 Jun.
Genetics v. 131 (2): p. 287-296; 1992 Jun. Includes
references.
Language: English
Descriptors: Endomycetales; Loci; Mating; Gene expression; Controlling elements; Genes; Genetic regulation; Transcription; Messenger RNA; Mutants; Meiosis; Recombination; Gene mapping; Gene location; Induced mutations; Semidominance
Abstract: The mat2-P and mat3-M loci of fission yeast contain respectively the plus (P) and minus (M) mating-type information in a transcriptionally silent state. That information is transposed from the mat2 or mat3 donor locus via recombination into the expressed mating-type locus (mat1) resulting in switching of the cellular mating type. We have identified a gene, named clr1 (for cryptic loci regulator), whose mutations allow expression of the mat2 and mat3 loci. clr1 mutants undergo aberrant haploid meiosis, indicative of transcription of the silent genes. Production of mRNA from mat3 is detectable in ch1 mutants. Furthermore, the ura4 gene inserted near mat3, weakly expressed in wild-type cells, is derepressed in clr1 mutants. The clr1 mutations also permit meiotic recombination in the 15-kb mat2-mat3 interval, where recombination is normally inhibited. The clr1 locus is in the right arm of chromosome II. We suggest that clr1 regulates silencing of the mat2 and mat3 loci, and participates in establishing the 'cold spot' for recombination by organizing the chromatin structure of the mating-type region.
26 NAL Call. No.: 442.8 Z34 Construction of a fusion gene comprising the Taka-amylase A promoter and the Escherichia coli beta-glucuronidase gene and analysis of its expression in Aspergillus oryzae. Tada, S.; Gomi, K.; Kitamoto, K.; Takahashi, K.; Tamura, G.; Hara, S. Berlin, W. Ger. : Springer International; 1991 Oct. M G G : Molecular and general genetics v. 229 (2): p. 301-306; 1991 Oct. Includes references.
Language: English
Descriptors: Aspergillus oryzae; Escherichia coli; Genetic engineering; Recombinant DNA; Promoters; Alpha-amylase; Reporter genes; Beta-glucuronidase; Gene expression; Messenger RNA; Starch; Sugars; Carbohydrate metabolism; Genetic transformation
Abstract: Northern blot analysis of glucose-grown and starchgrown mycelia of Aspergillus oryzae RIB40 was conducted using the cloned Taka-amylase A (TAA) gene as a probe. The amount of mRNA homologous to the TAA gene was increased when this fungus was grown with starch as a sole carbon source. In order to analyze the induction mechanism, we inserted the Escherichia coli uidA gene encoding beta-glucuronidase (GUS) downstream of the TAA promoter and introduced the resultant fusion gene into the A. oryzae genome. Production of a functional GUS protein was induced by starch, but not by glucose. When the effects of various sugars on expression of the fusion gene were examined, the results suggested that the expression of the fusion gene was under control of the TAA gene promoter.
27 NAL Call. No.: QH450.C74
Control of gene expression and the yeast cell cycle.
Wittenberg, C.; Reed, S.I.
Boca Raton, Fla. : CRC Press, Inc; 1991.
Critical reviews in eukaryotic gene expression v. 1 (3): p.
189-205; 1991. Literature review. Includes references.
Language: English
Descriptors: Saccharomyces cerevisiae; Gene expression; Genetic regulation; Cell division; Transcription; Literature reviews
28 NAL Call. No.: 472 N21 Coordination of expression of DNA synthesis genes in budding yeast by a cell-cycle regulated trans factor. Lowndes, N.F.; Johnson, A.L.; Johnston, L.H. London : Macmillan Magazines Ltd; 1991 Mar21. Nature v. 350 (6315): p. 247-250. ill; 1991 Mar21. Includes references.
Language: English
Descriptors: Saccharomyces; Gene expression; Genes; Dna; Synthesis
Abstract: All of the DNA synthesis genes of budding yeast examined so far are periodically expressed and hence under cell-cycle control. Expression occurs near the G1/S phase boundary and the genes seem to be coordinately regulated. The upstream promoter sequences of these genes have only a hexamer element, ACGCGT (an MluI restriction site), in common. Here we show that this hexamer is able to impart periodic expression to a heterologous gene and, significantly, this expression occurs coincidentally with that of CDC9, one of the DNA synthesis genes. We have also identified a protein that binds specifically to these sequences in a similar periodic manner. These ACGCGT sequences and the transcription factor that binds to them therefore seem to be the elements controlling both the periodic expression and coordinate regulation of the DNA synthesis genes.
29 NAL Call. No.: 500 N21P
Cotranslational autoproteolysis involved in gene expression
from a double-stranded RNA genetic element associated with
hypovirulence of the chestnut blight fungus.
Choi, G.H.; Shapira, R.; Nuss, D.L.
Washington, D.C. : The Academy; 1991 Feb15.
Proceedings of the National Academy of Sciences of the United
States of America v. 88 (4): p. 1167-1171; 1991 Feb15.
Includes references.
Language: English
Descriptors: Cryphonectria parasitica; Fungal diseases; Gene expression; Genetic control; Hypovirulence; Nucleotide sequences; Proteolysis; Translation
30 NAL Call. No.: 442.8 Z34 CYP1 (HAP1) is a determinant effector of alternative expression of heme-dependent transcription in yeast. Verdiere, J.; Gaisne, M.; Labbe-Bois, R. Berlin, W. Ger. : Springer International; 1991 Aug. M G G : Molecular and general genetics v. 228 (1/2): p. 300-306; 1991 Aug. Includes references.
Language: English
Descriptors: Saccharomyces cerevisiae; Controlling elements; Dna binding proteins; Genetic regulation; Gene expression; Genes; Oxidoreductases; Cytochrome p-450; Transcription; Anaerobic conditions; Oxygen; Heme; Deficiency
Abstract: The CYP1 (HAP1) gene of Saccharomyces cerevisiae is known to activate a number of target genes in response to the presence of heme. Several features of the protein, deduced from the sequence of the gene, suggest that CYP1 is a general sensor of the redox state of the cell. To investigate further the function of CYP1, we analysed its effects on the transcription of two genes, HEM13 and 14DM, which are preferentially expressed in anaerobiosis. HEM13 encodes coproporphyrinogen oxidase which catalyses the sixth enzymatic step in the heme biosynthetic pathway and 14DM encodes lanosterol-14-demethylase which is involved in sterol biosynthesis and is a member of the cytochrome P450 family. Isogenic CYP1+ and cyp1 deleted strains, either hemesufficient or heme-deficient (HEM1 disrupted), were grown in aerobic or anaerobic conditions, and transcripts of HEM13 and 14DM were analysed on Northern blots. The results show that in anaerobic and in heme-deficient cells, CYP1 activates the transcription of HEM13 and inhibits that of 14DM. Opposite effects of CYP1 are observed in aerobic heme-sufficient cells. We concluded that: (i) CYP1 is an efficient activator especially in heme-depleted cells; (ii) CYP1 exerts both positive and negative regulatory effects; (iii) the nature of the regulatory function of CYP1 depends on the target gene; and (iv) for a given gene, the presence or absence of heme or oxygen reverses the sense of CYP1-dependent regulation.
31 NAL Call. No.: 448.3 J82
Developmental regulation of CUP gene expression through DNA
methylation in Mucor spp.
Cano-Canchola, C.; Sosa, L.; Fonzi, W.; Sypherd, P.; RuizHerrera,
J. Washington, D.C. : American Society for
Microbiology; 1992 Jan. Journal of bacteriology v. 174 (2): p.
362-366; 1992 Jan. Includes references.
Language: English
Descriptors: Mucor; Genes; Gene expression; Dna; Methylation; Developmental stages; Saccharomyces cerevisiae; Transcription
Abstract: Inserts which carried the CUP gene from Saccharomyces cerevisiae or Mucor racemosus were used as hybridization probes to measure the methylation state and expression of the CUP gene from Mucor rouxii at different stages of growth. It was observed that the fungus contains a CUP multigene family. All the CUP genes were present in a hypermethylated DNA region in nongrowing and isodiametrically growing spores and were not transcribed at these stages. After germ tube emergence, CUP genes became demethylated and transcriptionally active. Development, demethylation, and transcription of CUP genes were blocked by the ornithine decarboxylase inhibitor 1,4-diaminobutanone. These results suggest that genes that are activated during development become demethylated in this fungus.
32 NAL Call. No.: QH442.A1G4
Dictyostelium discoideum as an expression host for the
circumsporozoite protein of Plasmodium falciparum.
Fasel, N.; Begdadi-Rais, C.; Bernard, M.; Bron, C.; Corradin,
G.; Reymond, C.D.
Amsterdam : Elsevier Science Publishers; 1992.
Gene v. 111 (2): p. 157-163; 1992. Includes references.
Language: English
Descriptors: Dictyostelium; Plasmodium falciparum; Cloning; Vectors; Genes; Antigens; Recombinant DNA; Gene expression; Promoters; Recombinant vaccines; Proteins
Abstract: We have used the cellular slime mold, Dictyostelium discoideum (Dd), to express the Plasmodium falciparum circumsporozoite protein (CS), a potential component of a subunit vaccine against malaria. This was accomplished via an expression vector based on the discoidin I-encoding gene promoter, in which we linked a sequence coding for a Dd leader peptide to the almost complete CS coding region (pEDII-CS). CS production at both the mRNA and protein levels is induced by starving cells in a simple phosphate buffer. Variation in pH or cell density does not seem to influence CS synthesis. CSproducing cells can be grown either on their normal substrate, bacteria, or on a semi-synthetic media, without affecting CS accumulation level. The CS produced in Dd seems similar to the natural parasite protein as judged by its size and epitope recognition by a panel of monoclonal antibodies. We constructed a second expression vector in which the CS is under the control of a Dd ras promoter. CS accumulation can then be induced by external addition of cAMP. Such a tightly regulated promoter may allow expression of proteins potentially toxic to the cell. Thus, Dd could be a useful eukaryotic system to produce recombinant proteins, in particular from human or animal parasites like P. falciparum.
33 NAL Call. No.: 448.3 J823 Differential gene expression and evidence of selective translation during anaerobic germination of Mucor racemosus sporangiospores. Linz, J.E.; Orlowski, M. Reading : Society for General Microbiology; 1991 Apr. The Journal of general microbiology v. 137 (pt.4): p. 827-835; 1991 Apr. Includes references.
Language: English
Descriptors: Mucor; Sporangia; Spores; Spore germination; Protein synthesis; Messenger RNA; Gene expression; Kinetics
Abstract: Anaerobically-germinated sporangiospores of Mucor racemosus develop into yeasts, whereas aerobically-germinated sporangiospores become hyphae. Anaerobic germination was found to have the following traits in common with the previously characterized aerobic system of development: (i) immediate and vigorous protein synthesis upon exposure of the spores to liquid medium; (ii) a complete absence of RNA synthesis for the first 20 min of germination; and (iii) a mobilization of ribosomal subunits into active polyribosomes. The proteins synthesized in both systems during this 20 min interval must be specified by the pre-formed stable mRNA known to be stored in the dormant spore. The population of proteins manufactured early in anaerobic germination differed considerably from the set of proteins synthesized during the equivalent interval in air, suggesting that some unknown mechanism of selective translation must operate. A few dozen of the more prominent proteins could be categorized according to their patterns of synthesis during germination. This should allow future work to focus upon those genes and their products most closely linked to development. The most promising candidates include several proteins that are most conspicuous in the mature yeast and are among those proteins selectively translated in large amounts from stored mRNA templates during anaerobic germination.
34 NAL Call. No.: QD341.A2N8
Direct introduction and transient expression of capped and
non-capped RNA in Saccharomyces cerevisiae.
Russell, P.J.; Hambidge, S.J.; Kirkegaard, K.
Oxford : IRL Press; 1991 Sep25.
Nucleic acids research v. 19 (18): p. 4949-4953; 1991 Sep25.
Includes references.
Language: English
Descriptors: Saccharomyces cerevisiae; Messenger RNA; Vectors; Translocation
Abstract: We report the introduction of functional RNA molecules into yeast spheroplasts. Plasmids containing the firefly luciferase coding region were transcribed to yield RNAs suitable for introduction into yeast cells and direct assay of their translation products. The 5' noncoding regions of the RNAs were derived either from the 5' noncoding regions of firefly luciferase, poliovirus, or yeast-like-particle (VLP) L-A or M1 RNAs. Capped and noncapped MRNAs were made by T7 RNA polymerase-directed transcription and introduced into yeast spheroplasts. The peak time of luciferase transient expression from introduced RNAs was 2 - 4 h after their introduction. in contrast, transient expression of luciferase from a non-replicative, luciferase-encoding plasmid introduced into the cells was maximal at 16 h. For capped mRNAs, luciferase activity increased linearly with transcript amount for both yeast and human (HeLa) cells. Although non-capped luciferase mRNAs were expressed more efficiently following introduction into yeast than into HeLa cells, the 5' noncoding sequences from yeast double-stranded (dS)RNA VLP RNAs conferred no greater apparent cap-independence than non-VLP RNA sequences in this transient expression assay. The RNA transient expression system will allow the study of translation of capped and non-capped RNAs in yeast cells and of the replicative cycle of yeast virus-like RNA genomes.
35 NAL Call. No.: 442.8 J828 Domain structure in actin-binding proteins: expression and functional characterization of truncated severin. Eichinger, L.; Noegel, A.A.; Schleicher, M. New York, N.Y. : Rockefeller University Press; 1991 Feb. The Journal of cell biology v. 112 (4): p. 665-676; 1991 Feb. Includes references.
Language: English
Descriptors: Fungi; Protein composition; Actin; Plant proteins; Mutants; Gene expression; Protein synthesis; Binding site; Genetic code
Abstract: Severin from Dictyostelium discoideum is a Ca2+- activated actin-binding protein that severs actin filaments, nucleates actin assembly, and caps the fast growing ends of actin filaments. Sequence comparison with functionally related proteins, such as gelsolin, villin, or fragmin revealed highly conserved domains which are thought to be of functional significance. To attribute the different activities of the severin molecule to defined regions, progressively truncated severin polypeptides were constructed. The complete cDNA coding for 362 (DS362) amino acids and five 3' deletions coding for 277 (DS277), 177 (DS177), 151 (DS151), 117 (DS117), or 111 (DS111) amino acids were expressed in Escherichia coli. The proteins were purified to homogeneity and then characterized with respect to their effects on the polymerization or depolymerization kinetics of G- or F-actin solutions and their binding to G-actin. Furthermore, the Ca2+ binding of these proteins was investigated with a 45Ca-overlay assay and by monitoring Ca2+-dependent changes in tryptophan fluorescence. Bacterially expressed DS362 showed the same Ca2+-dependent activities as native severin. DS277, missing the 85 COOH-terminal amino acids of severin, had lost its strict Ca2+ regulation and displayed a Ca2+-independent capping activity, but was still Ca2+ dependent in its severing and nucleating activities. DS151 which corresponded to the first domain of gelsolin or villin had completely lost severing and nucleating properties. However, a residual severing activity of approximately 2% was detectable if 26 amino acids more were present at the COOH-terminal end (DS177). This locates similar to gelsolin the second actinbinding site to the border region between the first and second domain. Measuring the fluorescence enhancement of pyrenelabeled G-actin in the presence of DS111 showed that the first actin-binding site was present in the NH2-terminal 111 amino acids. Extension by six or more amino acids stabilized t
36 NAL Call. No.: 385 J822 A dominant mutation that alters the regulation of INO1 expression in Saccharomyces cerevisiae. Hosaka, K.; Nikawa, J.; Kodaki, T.; Yamashita, S. Tokyo : Japanese Biochemical Society; 1992 Mar. Journal of biochemistry v. 111 (3): p. 352-358; 1992 Mar. Includes references.
Language: English
Descriptors: Saccharomyces cerevisiae; Genes; Mutations; Gene expression; Genetic regulation; Choline; Myo-inositol; Biosynthesis
Abstract: A dominant, single nuclear gene mutation, CSE1, caused inositol auxotrophy in yeast cells. The inositol requirement was marked when choline was present in the medium. Inositol-1-phosphate synthase, the regulatory enzyme of inositol synthesis, is repressed by inositol, or more profoundly by a combination of inositol and choline in the wild type. In CSE1, the level of inositol-1-phosphate synthase was low and was greatly repressed on the addition of choline alone. In accordance with this, INO1 mRNA encoding the enzyme was low even under the derepressed conditions and was profoundly decreased by choline in CSE1. But in the wild type, the addition of choline alone had little effect. An INO1-lacZ fusion was constructed and the control of the INO1 promoter in CSE1 was studied. lacZ expression was repressed not only by inositol, but also by choline in CSE1, whereas it was repressed by inositol, but only slightly by choline in the wild type. CSE1 was unlinked to the INO1 structural gene. Thus CSE1 was thought to be a regulatory mutation. Furthermore, when the CDP-choline pathway was mutationally blocked, choline did not affect IN01 expression, indicating that the metabolism of choline via the CDP-choline pathway is required for INO1 repression.
37 NAL Call. No.: 23 AU792
Effect of Kabatiella caulivora isolates and host growth stage
on symptom expression and resistance in Trifolium
subterraneum.
Barbetti, M.J.; Gillespie, D.J.; Collins, W.J.
East Melbourne : Commonwealth Scientific and Industrial
Research Organization; 1991.
Australian journal of experimental agriculture v. 31 (1): p.
63-69; 1991. Includes references.
Language: English
Descriptors: Western australia; Trifolium subterraneum; Cultivars; Crop growth stage; Disease resistance; Screening; Susceptibility; Symptoms; Crop damage; Kabatiella caulivora; Outbreaks; Pathogenicity
38 NAL Call. No.: QP501.E8 Efficient expression of bovine alpha-lactalbumin in Saccharomyces cerevisiae. Viaene, A.; Vvolckaert, G.; Joniau, M.; Baetselier, A. de; Cauwelaert, F. van New York, NY : Springer-Verlag New York Inc; 1991 Dec. European journal of biochemistry v. 202 (2): p. 471-477; 1991 Dec. Includes references.
Language: English
Descriptors: Saccharomyces cerevisiae; Alpha-lactalbumin; Synthetic genes; Recombination; Genetic transformation; Protein secretion; Gene expression; Ph; Amino acid sequences; Nucleotide sequences; Characterization
Abstract: A synthetic gene encoding the mature bovine alphalactalbumin fused to the preproregion of the yeast alphamating factor has been expressed and secreted at high level in Saccharomyces cerevisiae under the control of the alpha-mating promotor. Growth conditions were found to be critical for the expression: recombinant alpha-lactalbumin could only be detected in the medium provided the culture was grown at neutral pH. The secreted bovine alpha-lactalbumin is enzymatically active and identical to the whey protein, as confirmed by SDS/PAGE, IEF, ultraviolet and CD spectral analysis, and amino-terminal sequence determination.
39 NAL Call. No.: QH324.C7
Enhanced survival of yeast expressing an antifreeze gene
analogue after freezing.
McKown, R.L.; Warren, G.J.
Orlando, Fla. : Academic Press; 1991 Oct.
Cryobiology v. 28 (5): p. 474-482; 1991 Oct. Includes
references.
Language: English
Descriptors: Saccharomyces cerevisiae; Bacterial proteins; Cold resistance; Cryoprotectants; Freezing; Gene expression; Genetic transformation; Nucleotide sequences; Survival
40 NAL Call. No.: QH442.B5
Evaluation of foreign gene codon optimization in yeast:
expression of a mouse Ig kappa chain.
Kotula, L.; Curtis, P.J.
New York, N.Y. : Nature Publishing Company; 1991 Dec.
Bio/technology v. 9 (12): p. 1386-1389; 1991 Dec. Includes
references.
Language: English
Descriptors: Saccharomyces cerevisiae; Genetic engineering; Genetic transformation; Gene expression; Immunoglobulin structural genes; Mice; Genetic code; Protein synthesis
41 NAL Call. No.: 442.8 J828
Evolutionary conservation of the human nucleolar protein
fibrillarin and its functional expression in yeast.
Jansen, R.P.; Hurt, E.C.; Kern, H.; Lehtonen, H.; CarmoFonseca,
M.; Lapeyre, B.; Tollervey, D.
New York, N.Y. : Rockefeller University Press; 1991 May.
The Journal of cell biology v. 113 (4): p. 715-729; 1991 May.
Includes references.
Language: English
Descriptors: Yeasts; Protein synthesis; Genetic code; Nucleotide sequences; Amino acid sequences; Cell ultrastructure
Abstract: NOP1 is an essential nucleolar protein in yeast that is associated with small nucleolar RNA and required for ribosome biogenesis. We have cloned the human nucleolar protein, fibrillarin, from a HeLa cDNA library. Human fibrillarin is 70% identical to yeast NOP1 and is also the functional homologue since either human or Xenopus fibrillarin can complement a yeast nop1(-) mutant. Human fibrillarin is localized in the yeast nucleolus and associates with yeast small nucleolar RNAs. This shows that the signals within eucaryotic fibrillarin required for nucleolar association and nucleolar function are conserved from yeast to man. However, human fibrillarin only partially complements in yeast resulting in a temperature-sensitive growth, concomitantly altered rRNA processing and aberrant nuclear morphology. A suppressor of the human fibrillarin ts-mutant was isolated and found to map intragenically at a single amino acid position of the human nucleolar protein. The growth rate of yeast nop1(-) strains expressing Xenopus or human fibrillarin or the human fibrillarin suppressor correlates closely with their ability to efficiently and correctly process pre-rRNA. These findings demonstrate for the first time that vertebrate fibrillarin functions in ribosomal RNA processing in vivo.
42 NAL Call. No.: 381 J824
Expression and characterization of a recombinant yeast
isoleucyl-tRNA synthetase.
Racher, K.I.; Kalmar, G.B.; Borgford, T.J.
Baltimore, Md. : American Society for Biochemistry and
Molecular Biology; 1991 Sep15.
The Journal of biological chemistry v. 266 (26): p.
17158-17164; 1991 Sep15. Includes references.
Language: English
Descriptors: Saccharomyces cerevisiae; Isoleucine; Transfer RNA; Ligases; Recombinant DNA; Cloning; Gene expression; Enzyme activity; Kinetics
Abstract: We describe the heterologous expression of a recombinant Saccharomyces cerevisiae isoleucyl-tRNA synthetase (IRS) gene in Escherchia coli, as well as the purification and characterization of the recombinant gene product. High level expression of the yeast isoleucyl-tRNA synthetase gene was facilitated by site-specific mutagenesis. The putative ribosome-binding site of the yeast IRS gene was made to be the consensus of many highly expressed genes of E. coli. Mutagenesis simultaneously created a unique BclI restriction site such that the gene coding region could be conveniently subcloned as a "cassette." The variant gene was cloned into the expression vector pKK223-3 (Brosius, J., and Holy, A. (1984) Proc. Natl. Acad. Sci. U.S.A. 81, 6929-6933) thereby creating the plasmid pKR4 in which yeast IRS expression is under the control of the isopropyl-thio-beta-galactopyranoside (IPTG)-inducible tac promoter. Recombinant yeast IRS, on the order of 10 mg/liter of cell culture, was purified from pKR4- infected and IPTG-induced E. coli strain TG2. Yeast IRS was purified to homogeneity by a combination of anion-exchange and hydroxyapatite gel chromatography. Inhibition of yeast IRS activity by the antibiotic pseudomonic acid A was tested. The yeast IRS enzyme was found to be 10(4) times less sensitive to inhibition by pseudomonic acid A (Ki = 1.5 X 10(-5) M) than the E. coli enzyme. E. coli strain TG2 infected with pKR4, and induced with IPTG, had a plating efficiency of 100% at inhibitor concentrations in excess of 25 micromole/ml. At the same concentration of pseudomonic acid A, E. coli strain TG2 infected with pKK223-3 had a plating efficiency < 1%. The ability of yeast IRS to rescue E. coli from pseudomonic acid A suggests that the eukaryotic synthetase has full activity in its prokaryotic host and has specificity for E. coli tRNAile.
43 NAL Call. No.: 381 AR2
Expression and function of heterologous forms of malate
dehydrogenase in yeast.
Steffan, J.S.; Minard, K.I.; McAlister-Henn, L.
Orlando, Fla. : Academic Press; 1992 Feb14.
Archives of biochemistry and biophysics v. 293 (1): p. 93-102;
1992 Feb14. Includes references.
Language: English
Descriptors: Saccharomyces cerevisiae; Vectors; Mitochondria; Malate dehydrogenase; Gene expression; Genetic transformation; Enzyme activity; Escherichia coli; Rats
44 NAL Call. No.: QP251.I628 Expression and secretion in yeast of active insect defensin, an inducible antibacterial peptide from the fleshfly Phormia terranovae. Reichhart, J.M.; Petit, I.; Legrain, M.; Dimarcq, J.L.; Keppi, E.; Lecocoq, J.P.; Hoffmann, J.A.; Achstetter, T. Rehovot, Israel : Balaban Publishers; 1992 Feb. Invertebrate reproduction & development v. 21 (1): p. 15-24; 1992 Feb. Includes references.
Language: English
Descriptors: Phormia terraenovae; Peptides; Antibacterial properties; Gene expression; Saccharomyces cerevisiae; Nucleotide sequences; Amino acid sequences
45 NAL Call. No.: 448.3 J823
Expression and secretion of staphylococcal nuclease in yeast:
effects of amino-terminal sequences.
Pines, O.; London, A.
Reading : Society for General Microbiology; 1991 Apr.
The Journal of general microbiology v. 137 (pt.4): p. 771-778;
1991 Apr. Includes references.
Language: English
Descriptors: Saccharomyces cerevisiae; Staphylococcus aureus; Nucleases; Enzyme activity; Amino acid sequences; Protein secretion; Genes; Gene expression; Clones
Abstract: Staphylococcus aureus nuclease A hybrid genes, encoding proteins OmpA-nuclease, lipo-nuclease and Pinnuclease, were cloned downstream of the yeast GAL1O inducible promoter. OmpA-nuclease and lipo-nuclease contain the mature staphylococcal nuclease sequence preceded by the Escherichia coli OmpA and lipoprotein signal sequences, respectively, whereas Pin-nuclease lacks a defined signal sequence at its amino terminus. We found that: (a) the nuclease gene products synthesized in yeast are active, but they do not affect cell growth; b) OmpA-nuclease and lipo-nuclease are partially processed and constitute approximately 1.0-1.5% of the yeast cell protein; (c) OmpA and lipoprotein signal sequences function similarly in secretion, allowing 35-40% of the processed nuclease to be translocated into the yeast periplasm; and (d) Pin-nuclease, which lacks hydrophobic sequences at its amino-terminus, is accumulated at a level tenfold lower than the hybrid proteins that do contain signal sequences. Nevertheless, 50% of the enzyme activity of Pinnuclease in yeast is localized in the periplasmic space.
46 NAL Call. No.: QH426.C8
Expression and sequence comparison of the Aspergillus niger
and Aspergillus tubigensis genes encoding polygalacturonase
II.
Bussink, H.J.D.; Buxton, F.P.; Visser, J.
Berlin, W. Ger. : Springer International; 1991.
Current genetics v. 19 (6): p. 467-474; 1991. Includes
references.
Language: English
Descriptors: Aspergillus niger; Aspergillus; Gene expression; Genetic code; Polygalacturonase; Nucleotide sequences; Restriction mapping; Amino acid sequences
Abstract: The structure and expression of the polygalacturonase-encoding pgaII genes of two recently recognized species, Aspergillus niger and Aspergillus tubigensis, was investigated. While the structure of the pgaII genes is very similar, showing 83% DNA sequence identity and 94% identity at the amino acid level, they have diverged significantly. The NH2-terminal sequence suggests that these PGs are made as pre pro-proteins and the secretory propeptide of the PGII precursors shows sequence homology with some other fungal pro-peptides. The expression of the pgaII genes is strongly regulated by the carbon source and the A. tubigensis gene is expressed and regulated in A. niger transformants. The low similarity of the fungal PGs with those of bacterial and plant origin is discussed in relation to the possible functional role of specific amino acids.
47 NAL Call. No.: 448.3 J82 Expression in Escherichia coli of the Saccharomyces cerevisiae CCT gene encoding cholinephosphate cytidylyltransferase. Tsukagoshi, Y.; Nikawa, J.; Hosaka, K.; Yamashita, S. Washington, D.C. : American Society for Microbiology; 1991 Mar. Journal of bacteriology v. 173 (6): p. 2134-2136. ill; 1991 Mar. Includes references.
Language: English
Descriptors: Saccharomyces cerevisiae; Transferases; Genetic code; Cloning; Plasmids; Escherichia coli; Enzyme activity; Gene expression
Abstract: The coding region of the CCT gene from the yeast
Saccharomyces cerevisiae was cloned into the pUC18 expression
vector. The plasmid directed the synthesis of an active
cholinephosphate cytidylyltransferase in Escherichia coli,
confirming that CCT is the structural gene for this enzyme.
The enzyme produced in E. coli efficiently utilized
cholinephosphate and N,N-dimethylethanolaminephosphate, but Nmethylethanolaminephosphate
and ethanolaminephosphate were
poor substrates. Consistently, disruption of the CCT locus in
the wild-type yeast cells resulted in a drastic decrease in
activities with respect to the former two substrates. When
activity was expressed in E. coli, over 90% was recovered in
the cytosol, whereas most of the activity of yeast cells was
associated with membranes, suggesting that yeast cells possess
a mechanism that promotes membrane association of
cytidylyltransferase.
48 NAL Call. No.: 442.8 Z34
Expression in yeast of a cDNA copy of the K2 killer toxin
gene. Dignard, D.; Whiteway, M.; Germain, D.; Tessier, D.;
Thomas, D.Y. Berlin, W. Ger. : Springer International; 1991
May.
M G G : Molecular and general genetics v. 227 (1): p. 127-136;
1991 May. Includes references.
Language: English
Descriptors: Saccharomyces cerevisiae; Rna; Mycotoxins; Genes; Cloning; Nucleotide sequences; Amino acid sequences; Phytotoxicity; Resistance; Targeted mutagenesis; Induced mutations; Mutants; Enzyme activity; Proteinases; Gene expression
Abstract: A cDNA copy of the M2 dsRNA encoding the K2 killer toxin of Saccharomyces cerevisiae was expressed in yeast using the yeast ADH1 promoter. This construct produced K2-specific killing and immunity functions. Efficient K2-specific killing was dependent on the action of the KEX2 endopeptidase and the KEX1 carboxypeptidase, while K2-specific immunity was independent of these proteases. Comparison of the K2 toxin sequence with that of the K1 toxin sequence shows that although they share a common processing pathway and are both encoded by cytoplasmic dsRNAs of similar basic structure, the two toxins are very different at the primary sequence level. Site-specific mutagenesis of the cDNA gene establishes that one of the two potential KEX2 cleavage sites is critical for toxin action but not for immunity. Immunity was reduced by an insertion of two amino acids in the hydrophobic amino-terminal region which left toxin activity intact, indicating an independence of toxin action and immunity.
49 NAL Call. No.: QP501.B64
Expression of a cDNA clone encoding the haem-binding domain of
Chlorella nitrate reductase.
Cannons, A.C.; Iida, N.; Solomonson, L.P.
London : The Biochemical Society; 1991 Aug15.
The Biochemical journal v. 278 (pt.1): p. 203-209; 1991 Aug15.
Includes references.
Language: English
Descriptors: Chlorella vulgaris; Clones; Nitrate reductase; Heme; Binding site; Amino acid sequences; Nucleotide sequences; Spectral analysis
Abstract: A partial cDNA clone coding for the haem-binding domain of NADH:nitrate reductase (EC 1.6.6.1) (NR) from the unicellular green alga Chlorella vulgaris has been isolated, sequenced and expressed. A 1.2 kb cDNA (pCVNR1) was isolated from a lambda gt 11 expression library produced from polyadenylated RNA extracted from nitrate-grown Chlorella cells. pCVNR1 hybridized to a 3.5 kb mRNA transcript that was nitrate-inducible and absent from ammonium-grown cells. The entire sequence of pCVNR1 was obtained and found to have a single uninterrupted reading frame. The derived amino acid sequence of 318 amino acids has a 45-50 % similarity to higher-plant NRs, including Arabidopsis thaliana, spinach (Spinacia oleracea) and tobacco (Nicotiana tabacum). A comparison with the putative domain structure of higher-plant nitrate reductases suggested that this sequence contains the complete haem-binding domain, approximately one-third of the Mo-pterin domain and no FAD-binding domain. A 32% sequence similarity is evident when comparing the Chlorella NR haem domain with that of calf cytochrome b5. Expression of pCVNR1 in a pET vector synthesized a 35 kDa protein that was antigenic to anti-(Chlorella NR) antibody. The spectral properties of this protein (reduced and oxidized) in the 400-600 nm region are identical with those of native Chlorella NR and indicate that haem is associated with the protein.
50 NAL Call. No.: 442.8 Z34
Expression of a dominant negative allele of cdc2 prevents
activation of the endogenous p34cdc2 kinase.
Fleig, U.N.; Nurse, P.
Berlin, W. Ger. : Springer International; 1991 May.
M G G : Molecular and general genetics v. 226 (3): p. 432-440;
1991 May. Includes references.
Language: English
Descriptors: Endomycetales; Mutants; Alleles; Dominant lethals; Protein kinase; Serine; Enzyme activity; Cell division; Phosphoproteins; Amino acid sequences; Nucleotide sequences; Induced mutations
Abstract: The cdc2 gene of the fission yeast Schizosaccharomyces pombe encodes a 34 kDa phosphoprotein with serine/threonine protein kinase activity that acts as the key component in regulation of the eukaryotic cell cycle. We used a repressible promoter fused to the cdc2 cDNA to isolate conditionally dominant negative mutants of cdc2. One of these mutants, DL5, is described in this paper. Overexpression of the mutant protein in a wild-type cdc2 background is lethal and confers cell cycle arrest with a typical cdc-phenotype. Sequencing of the mutant cdc2 gene revealed a single amino acid substitution in a region highly conserved in cdc2-like proteins. The mutant protein exhibits no protein kinase activity, but is able to bind a component(s) required for an active protein kinase complex and thereby prevents binding of this component(s) to the co-existing wild-type cdc2 protein. We also demonstrate that S. pombe p34(cdc2) contains no phosphoserine.
51 NAL Call. No.: 450 P692 Expression of a fungal sesquiterpene cyclase gene in transgenic tobacco. Hohn, T.M.; Ohlrogge, J.B. Rockville, Md. : American Society of Plant Physiologists; 1991 Sep. Plant physiology v. 97 (1): p. 460-462; 1991 Sep. Includes references.
Language: English
Descriptors: Nicotiana tabacum; Transgenics; Fusarium sporotrichioides; Gene expression; Biosynthesis; Sesquiterpenes; Ligases; Enzyme activity; Disease resistance
Abstract: The complete coding sequence for the trichodiene synthase gene from Fusarium sporotrichioides was introduced into tobacco (Nicotiana tabacum) under the regulation of the cauliflower mosaic virus 35S promoter. Expression of trichodiene synthase was demonstrated in the leaves of transformed plants. Leaf homogenates incubated with [3H]farnesyl pyrophosphate produced trichodiene as a major product. Trichodiene was detected in the leaves of a transformed plant at a level of 5 to 10 nanograms per gram fresh weight. The introduction of a fungal sesquiterpene cyclase gene into tobacco has resulted in the expression of an active enzyme and the accumulation of low levels of its sesquiterpenoid product.
52 NAL Call. No.: TX393.J6
Expression of a wheat gliadin protein in yeast (Saccharomyces
cerevisiae). Pratt, K.A.; Madgwick, P.J.; Shewry, P.R.
London : Academic Press; 1991 Nov.
Journal of cereal science v. 14 (3): p. 223-229; 1991 Nov.
Includes references.
Language: English
Descriptors: Wheat; Gliadin; Gene expression; Saccharomyces cerevisiae
Abstract: A full-length CDNA encoding a gamma-type gliadin from wheat was expressed in Saccharomyces cerevisiae. A yield of 1-2% of total cell protein was obtained, and the level of expression was not affected by using a protease-deficient strain of yeast. Analysis of the purified gamma-gliadin showed that cleavage of the signal peptide had occurred but that incorrect disulphide bond formation had resulted in a range of oligomers and polymers in addition to monomers.
53 NAL Call. No.: 448.3 J82 Expression of bacterial mercuric ion reductase in Saccharomyces cerevisiae. Rensing, C.; Kues, U.; Stahl, U.; Nies, D.H.; Friedrich, B. Washington, D.C. : American Society for Microbiology; 1992 Feb. Journal of bacteriology v. 174 (4): p. 1288-1292; 1992 Feb. Includes references.
Language: English
Descriptors: Saccharomyces cerevisiae; Genetic transformation; Gene expression; Oxidoreductases; Mercury; Ions; Resistance
Abstract: The gene merA coding for bacterial mercuric ion reductase was cloned under the control of the yeast promoter for alcohol dehydrogenase I in the yeast-Escherichia coli shuttle plasmid pADH040-2 and transformed into Saccharomyces cerevisiae AH22. The resulting transformant harbored stable copies of the merA-containing hybrid plasmid, displayed a fivefold increase in the MIC of mercuric chloride, and synthesized mercuric ion reductase activity.
54 NAL Call. No.: SB599.P45
Expression of disease resistance response genes in nearisogenic
pea cultivars following challenge by Fusarium
oxysporum race 1.
Hadwiger, L.A.; Chiang, C.C.; Horovitz, D.
London : Academic Press; 1992 Apr.
Physiological and molecular plant pathology v. 40 (4): p.
259-269; 1992 Apr. Includes references.
Language: English
Descriptors: Pisum sativum; Fusarium oxysporum f.sp. pisi; Disease resistance; Genetic resistance; Cultivars; Lines; Gene expression; Dominance; Genes; Varietal susceptibility; Races; Host parasite relationships
Abstract: Previous research on the pea/Fusarium oxysporum system has shown that the internal invasion of tap roots and hypocotyls was less frequent in the cultivar Vantage, which possesses a single dominant gene for resistance to Race 1, than in the cultivar M410 which is susceptible. We monitored these cultivar tissues for the expression of disease resistance response genes 49 and 206, previously associated with the expression of non-host resistance, in peas grown in soil naturally infested with Fusarium oxysporum f. sp. pisi (Race 1). The accumulation of distinct RNA products homologous with both genes was evident in the resistant cultivar in a period prior to and during the onset of wilt symptoms in the susceptible cultivar. Additional periods of RNA accumulation occurred in older flowering seedlings of both cultivars. These were not gene-specific RNA accumulations and were probably unrelated to resistance. Thus these response genes may contribute to normal cellular functions which are also utilized in the disease resistance response. The biochemical functions of genes 49 and 206 remain unknown.
55 NAL Call. No.: QK600.B72
Expression of heterologous genes in filamentous fungi.
Davies, R.W.
Cambridge : Cambridge University Press; 1991.
Symposium series - British Mycological Society (18): p.
103-117; 1991. In the series analytic: Applied molecular
genetics of fungi / edited by J. F. Peberdy, C. E. Caten, J.
E. Ogden and J. W. Bennett. Symposium of the British
Mycological Society held at the University of Nottingham,
April 1990. Literature review. Includes references.
Language: English
Descriptors: Fungi; Aspergillus nidulans; Aspergillus niger; Gene transfer; Recombinant DNA; Gene expression; Genetic transformation; Proteins; Structural genes; Vectors; Plasmids; Literature reviews
56 NAL Call. No.: QH442.A1G4
Expression of lacZ gene fusions affects downstream
transcription in yeast. Barnes, C.A.; Johnston, G.C.; Singer,
R.A.
Amsterdam : Elsevier Science Publishers; 1991.
Gene v. 104 (1): p. 47-54; 1991. Includes references.
Language: English
Descriptors: Saccharomyces cerevisiae; Genes; Gene expression; Transcription; Heat shock; Recombinant DNA
Abstract: Chimeric genes containing Escherichia coli lacZ sequences are often used to characterize gene expression in yeast cells. By Northern analysis, we found that such genes produce multiple transcripts due to inefficient 3'-end formation. The same transcript pattern was found for two related chimeric genes when these genes were cloned separately into the commonly used vector, YIp5, and integrated into the yeast genome at two different locations. Each chimeric gene was composed of promoter and N-terminal coding regions from the yeast SSA1 or SSA2 genes fused in-frame to the lac operon. Transcripts were shown to initiate within the yeast promoter fragment, but transcript size indicated that 3' ends were localized to three different regions: within the lac operon near the 3' end of the lacZ gene; near a terminator region previously identified upstream of the URA3 gene in YIp5; and at the URA3 terminator region. Readthrough transcription of the URA3 promoter from upstream lac sequences decreased the basal activity of the URA3 promoter, although induced URA3 transcription levels were unaffected. This readthrough transcription also resulted in a novel, longer URA3 transcript.
57 NAL Call. No.: QR360.A1J6
Expression of potyvirus coat protein in Escherichia coli and
yeast and its assembly into virus-like particles.
Jagadish, M.N.; Ward, C.W.; Gough, K.H.; Tulloch, P.A.;
Whittaker, L.A.; Shukla, D.D.
Reading : Society for General Microbiology; 1991 Jul.
The Journal of general virology v. 72 (pt.7): p. 1543-1550;
1991 Jul. Includes references.
Language: English
Descriptors: Potyvirus group; Coat proteins; Escherichia coli; Saccharomyces cerevisiae
Abstract: When the full-length coat protein (CP) of the potyvirus, Johnsongrass mosaic virus (JGMV), was expressed in Escherichia coli or yeast, it assembled to form potyvirus-like particles. The particles were heterogeneous in length with a stacked-ring appearance and resembled JGMV particles in their flexuous morphology and width. This cell-free assembly system should permit analysis of the mechanisms of particle assembly and genome encapsidation. Two mutant forms of CP produced by site-directed mutagenesis failed to assemble into virus-like particles.
58 NAL Call. No.: 500 N21P
Expression of RNase P RNA in Saccharomyces cerevisiae is
controlled by an unusual RNA polymerase III promoter.
Lee, J.Y.; Evans, C.F.; Engelke, D.R.
Washington, D.C. : The Academy; 1991 Aug15.
Proceedings of the National Academy of Sciences of the United
States of America v. 88 (16): p. 6986-6990; 1991 Aug15.
Includes references.
Language: English
Descriptors: Saccharomyces cerevisiae; Strains; Gene expression; Genetic code; Nucleotide sequences; Promoters; Ribonucleases; Rna polymerase; Transcription
Abstract: The RNA subunit of Saccharomyces cerevisiae nuclear RNase P is encoded by a single-copy, essential gene, RPR1. The 369-nucleotide mature form of the RNA has an apparent precursor with an 84-nucleotide 5' leader and approximately 33 nucleotides of additional 3' sequence. Analysis of RPR1 transcription in a strain with a temperature-sensitive lesion in RNA polymerase III shows that the gene is transcribed in vivo by RNA polymerase III. Examination of potential promoter regions using both progressive upstream deletions and point mutations indicates that at least two sequences contained within the 5' leader region are essential for expression in vivo, while sequences farther upstream influence efficiency. The required leader elements resemble tRNA gene-like A-box and B-box internal promoters in sequence and spacing. As in the tRNA genes, transcription factor TFIIIC binds to this region in vitro and binding is severely reduced by either A-box or Bbox point mutations that impair expression in vivo. It thus appears that the yeast RNase P RNA gene has adopted a promoter strategy that places an RNA polymerase III "internal" promoter upstream of the mature structural domain to help drive transcription.
59 NAL Call. No.: 385 J822 Expression of RNase Rh from Rhizopus niveus in yeast and characterization of the secreted proteins. Ohgi, K.; Horiuchi, H.; Watanabe, H.; Takagi, M.; Yano, K.; Irie, M. Tokyo : Japanese Biochemical Society; 1991 May. Journal of biochemistry v. 109 (5): p. 776-785; 1991 May. Includes references.
Language: English
Descriptors: Rhizopus; Saccharomyces cerevisiae; Ribonucleases; Vectors; Gene expression; Dna; Nucleotide sequences; Amino acid sequences
60 NAL Call. No.: 448.3 AP5
Expression of the Escherichia coli beta-glucuronidase gene in
Pseudocercosporella herpotrichoides.
Bunkers, G.J.
Washington, D.C. : American Society for Microbiology; 1991
Oct. Applied and environmental microbiology v. 57 (10): p.
2896-2900; 1991 Oct. Includes references.
Language: English
Descriptors: Pseudocercosporella herpotrichoides; Genetic analysis; Genetic transformation; Escherichia coli; Betaglucuronidase; Enzyme activity; Gene expression; Genetic markers; Plant pathogens; Hordeum vulgare; Triticum aestivum; Secale cereale
Abstract: The plant-pathogenic fungus Pseudocercosporella herpotrichoides has been successfully transformed by using two different positive selection systems in combination with the Escherichia coli gusA gene. The selectable markers used in this study were the hygromycin B phosphotransferase gene (hph) from E. coli and the gene (bml) for beta-tubulin from a benomyl-resistant mutant of Neurospora crassa. A lower transformation rate was obtained with the bml system than with the hph system. Conversely, cotransformation frequencies, as determined with medium plates containing the chromogenic substrate 5-bromo-4-chloro-3-indolyl-beta-D-glucuronic acid, were higher with bml than with hph as the selectable marker. The hygromycin-resistant transformants were mitotically stable, and both the selectable gene and gusA were maintained through conidiation. The vector DNA was integrated into the genome, and the number and sites of insertion varied among transformants. Enzyme assays of mycelial extracts showed that beta-glucuronidase activity was highest in transformants with a high gusA copy number. Expression of gusA during growth of the fungus on plants was easily detectable and did not affect pathogenicity. These results form the basis for construction of a versatile and sensitive reporter gene system for P. herpotrichoides.
61 NAL Call. No.: 381 J824
Expression of the yeast plasma membrane [H+]ATPase in
secretory vesicles. A new strategy for directed mutagenesis.
Nakamoto, R.K.; Rao, R.; Slayman, C.W.
Baltimore, Md. : American Society for Biochemistry and
Molecular Biology; 1991 Apr25.
The Journal of biological chemistry v. 266 (12): p. 7940-7949;
1991 Apr25. Includes references.
Language: English
Descriptors: Saccharomyces cerevisiae; Plasma membranes; Adenosinetriphosphatase; Cytoplasmic inclusions; Secretory granules; Enzyme activity; Transport processes
Abstract: Secretory vesicles that accumulate in the temperature-sensitive sec6-4 strain of yeast have been shown to contain a vanadate-sensitive ATPase, presumably en route to the plasma membrane (Walworth, N. C., and Novick, P. J. (1987) J. Cell Biol. 105, 163-174). We have now established this enzyme to be a fully functional form of the PMA1 [H+]ATPase, identical in its catalytic properties to that found in the plasma membrane. In addition, the secretory vesicles are sealed tightly enough to permit the measurement of ATPdependent proton pumping with fluorescent probes. We have gone on to exploit the vesicles as an expression system for sitedirected mutants of the ATPase. For this purpose, a sec6-4 strain has been constructed in which the chromosomal PMA1 gene is under control of the GAL1 promoter; the mutant pma1 allele to be studied is introduced on a centromeric plasmid under the control of a novel heat shock promoter. In galactose medium at 23 degrees C, the wild-type ATPase is produced and supports normal vegetative growth. When the cells are switched to glucose medium at 37 degrees C, however, the wild-type gene turns off, the mutant gene turns on, and secretory vesicles accumulate. The vesicles contain a substantial amount of newly synthesized, plasmid-encoded ATPase (5-10% of total vesicle protein), but only traces of residual wild-type PMA1 ATPase and no detectable mitochondrial ATPase, vacuolar ATPase, or acid or alkaline phosphatase. To test the expression strategy, we have made use of pma1-105 (Ser 368 leads to Phe), a vanadate-resistant mutant previously characterized by standard methods (Perlin, D. S., Harris, S. L., Seto-Young, D., and Haber, J. E. (1989) J. Biol. Chem. 264, 21857-21864). In secretory vesicles, as expected, the plasmid-borne pma1 allele 105 gives rise to a mutant enzyme with a reduced rate of ATP hydrolysis and a 100-fold increase in Ki for vanadate. Proton pumping is similarly resistant to vanadate. Thus, the vesicles appear well suited f
62 NAL Call. No.: 500 N21P
Expression of three mammalian cDNAs that interfere with RAS
function in Saccharomyces cerevisiae.
Colicelli, J.; Nicolette, C.; Birchmeier, C.; Rodgers, L.;
Riggs, M.; Wigler, M.
Washington, D.C. : The Academy; 1991 Apr01.
Proceedings of the National Academy of Sciences of the United
States of America v. 88 (7): p. 2913-2917. ill; 1991 Apr01.
Includes references.
Language: English
Descriptors: Saccharomyces cerevisiae; Strains; Yeasts; Amino acid sequences; Cloning; Dna; Gene expression; Genetic code; Heat shock; Nucleotide sequences; Oncogenes
Abstract: Saccharomyces cerevisiae strains expressing the activated RAS2(Val19) gene or lacking both cAMP phosphodiesterase genes, PDE1 and PDE2, have impaired growth control and display an acute sensitivity to heat shock. We have isolated two classes of mammalian cDNAs from yeast expression libraries that suppress the heat shock-sensitive phenotype of a RAS2(Val19) strain. Members of the first class of cDNAs also suppress the heat shock-sensitive phenotype of pde1- pde2- strains and encode cAMP phosphodiesterases. Members of the second class fail to suppress the phenotype of pde1- pde2- strains and therefore are candidate cDNAs encoding proteins that interact with RAS proteins. We report the nucleotide sequence of three members of this class. Two of these cDNAs share considerable sequence similarity, but none are clearly similar to previously isolated genes.
63 NAL Call. No.: 442.8 Z34
Expression of variant nuclear Arabidopsis tRNA(Ser) genes and
pre-tRNA maturation differ in HeLa, yeast and wheat germ
extracts. Beier, D.; Beier, H.
Berlin, W. Ger. : Springer International; 1992 May.
M G G : Molecular and general genetics v. 233 (1/2): p.
201-208; 1992 May. Includes references.
Language: English
Descriptors: Arabidopsis thaliana; Transcription; Gene expression; Transfer RNA; Serine; Multiple genes; Precursors; Rna editing; Pseudogenes; In vitro; Extracts; Hela cells; Yeast extracts; Plant extracts; Wheat germ; Mutations; Messenger RNA; Nucleotide sequences; Molecular conformation; Restriction mapping
Abstract: We have recently identified a tRNA gene cluster in the Arabidopsis nuclear genome. One tRNA(Ser) (AGA) gene and two tRNA(Tyr) (GTA) genes occur in tandem arrangement on a 1.5 kb unit that is amplified about 20-fold at a single chromosomal site. Here we have studied the in vitro expression of seven individually cloned tRNA(Ser) genes (pAtS1 to pAtS7) derived from this cluster. Five out of the seven tRNA(Ser) genes contain point mutations in the coding region which have in part adverse effects on the expression of these genes in different cell-free systems: (i) C10 and A62 in plant tRNA(Ser) genes, which correspond to G10 and C62, respectively, in all known vertebrate tRNA genes, result in a reduced transcription efficiency in HeLa but not in yeast extract. This indicates that yeast RNA polymerase III tolerates nucleotide substitutions at positions 10 [5' internal control region (ICR)] and 62 (3' ICR), whereas the vertebrate RNA polymerase III requires a more stringent consensus sequence. (ii) Processing of a pre-tRNA(Ser) with a mismatch in the aminoacyl stem is impaired in HeLa, yeast and wheat germ extracts, however, a mismatch in the anticodon stem is deleterious for HeLa and wheat germ but not for yeast processing enzymes. The unexpectedly high number of potential tRNA(Ser) pseudogenes in the cluster--quite in contrast to the tRNA(Tyr) genes which mainly code for functional tRNAs-- suggested that tRNA(Ser) (AGA) genes also occur elsewhere in the genome. We present evidence that single copies of tRNA(Ser) (AGA) genes do indeed exist outside the tRNA gene cluster.
64 NAL Call. No.: 442.8 Z34 Expression of yeast cytochrome c1 is controlled at the transcriptional level by glucose, oxygen and haem. Oechsner, U.; Hermann, H.; Zollner, A.; Haid, A.; Bandlow, W. Berlin, W. Ger. : Springer International; 1992 Apr. M G G : Molecular and general genetics v. 232 (3): p. 447-459; 1992 Apr. Includes references.
Language: English
Descriptors: Saccharomyces cerevisiae; Structural genes; Cytochrome c; Gene expression; Genetic regulation; Glucose; Oxygen; Heme; Transcription; Promoters; Dna binding proteins; Binding site; Restriction mapping; Nucleotide sequences; Controlling elements
Abstract: The nuclear gene for cytochrome c1 in Saccharomyces cerevisiae (CYT1) was localized on chromosome XV. Its upstream region was identified by functional complementation. Fusion to the lacZ reporter gene on a CEN plasmid allowed study of the effect of carbon sources and of specific deletion mutations on expression of the gene in yeast transformants. Detailed promoter analysis combined with expression studies in recipient strains defective in regulatory genes identified cis-acting sites and transcription factors involved in the regulated expression of the cytochrome c1 gene. These analyses showed that, in the presence of glucose, transcription of CYT1 is positively controlled by oxygen, presumably through the haem signal, and mediated by the HAP1-encoded transactivator. It is additionally regulated by the HAP2/3/4 complex which mediates gene activation mainly under glucose-free conditions. Basal transcription is, in part, effected by CPF1, a centromere and promoter-binding factor.
65 NAL Call. No.: QR360.J6 Expression of yeast L-A double-stranded RNA virus proteins produces derepressed replication: a ski- phenocopy. Wickner, R.B.; Icho, T.; Fujimura, T.; Widner, W.R. Washington, D.C. : American Society for Microbiology; 1991 Jan. Journal of virology v. 65 (1): p. 155-161. ill; 1991 Jan. Includes references.
Language: English
Descriptors: Saccharomyces cerevisiae; Rna; Proteins; Replication
Abstract: The plus strand of the L-A double-stranded RNA virus of Saccharomyces cerevisiae has two large open reading frames, ORF1, which encodes the major coat protein, and ORF2, which encodes a single-stranded RNA-binding protein having a sequence diagnostic of viral RNA-dependent RNA polymerases. ORF2 is expressed only as a Gag-Pol-type fusion protein with ORF1. We have constructed a plasmid which expresses these proteins from the yeast PGK1 promoter. We show that this plasmid can support the replication of the killer toxinencoding M1, satellite virus in the absence of an L-A doublestranded RNA helper virus itself. This requires ORF2 expression, providing a potential in vivo assay for the RNA polymerase and single-stranded RNA-binding activities of the fusion protein determined by ORF2. ORF1 expression, like a host ski- mutation, can suppress the usual requirement of M1 for the MAK11, MAK18, and MAK27 genes and allow a defective LA (L-A-E) to support M1 replication. These results suggest that expression of ORF1 from the vector makes the cell a skiphenocopy. Indeed, expression of ORF1 in a wild-type killer makes it a superkiller, suggesting that a target of the SKI antiviral system may be the major coat protein.
66 NAL Call. No.: QH442.A1G4 An expression vector for the phytopathogenic fungus, Ustilago maydis. Kinal, H.; Tao, J.; Bruenn, J.A. Amsterdam : Elsevier Science Publishers; 1991. Gene v. 98 (1): p. 129-134; 1991. Includes references.
Language: English
Descriptors: Ustilago zeae; Vectors; Plasmids; Genetic transformation; Marker genes; Drug resistance; Hygromycin b; Dna; Cloning; Nucleotide sequences; Mycotoxins; Toxicity; Virus-like particles
Abstract: We have constructed an expression vector for the phytopathogenic fungus Ustilago maydis. This vector, pUXV, expresses genes located downstream from a U. maydis glyceraldehyde-3-phosphate dehydrogenase promoter. Plasmid pUXV also contains a selective marker gene conferring resistance to the antibiotic hygromycin B and a U. maydis autonomously replicating sequence, UARS, allowing high transformation efficiency. Expression of a cDNA from the toxin-encoding region of the U. maydis virus P6 in pUXV resulted in as much killing activity as from viral particles when evaluated by killer plate assay. Plasmid pUXV preserves essential sequences from pUC12 and is therefore a shuttle vector for U. maydis and Escherichia coli.
67 NAL Call. No.: 500 N21P Functional expression of a probable Arabidopsis thaliana potassium channel in Saccharomyces cerevisiae. Anderson, J.A.; Huprikar, S.S.; Kochian, L.V.; Lucas, W.J.; Gaber, R.F. Washington, D.C. : The Academy; 1992 May01. Proceedings of the National Academy of Sciences of the United States of America v. 89 (19): p. 3736-3740; 1992 May01. Includes references.
Language: English
Descriptors: Saccharomyces cerevisiae; Yeasts; Mutants; Potassium; Ion transport; Arabidopsis thaliana; Amino acid sequences; Cloning; Nucleotide sequences
Abstract: We report the isolation of a cDNA (KAT1) from Arabidopsis thaliana that encodes a probable K+ channel. KAT1 was cloned by its ability to suppress a K+ transport-defective phenotype in mutant Saccharomyces cerevisiae cells. This suppression is sensitive to known K+ channel blockers, including tetraethylammonium and Ba2+ ions. The KAT1 cDNA contains an open reading frame capable of encoding a 78-kDa protein that shares structural features found in the Shaker superfamily of K+ channels. These include a cluster of six putative membrane-spanning helices (S1-S6) at the amino terminus of the protein, a presumed voltage-sensing region containing Arg/Lys-Xaa-Xaa-Arg/Lys repeats within S4, and the highly conserved pore-forming region (known as H5 or SS1-SS2). Our results suggest that the structural motif for K+ channels has been conserved between plants and animals.
68 NAL Call. No.: 381 AR2
Functional expression of mammalian cytochromes P450IIB in the
yeast Saccharomyces cerevisiae.
Kedzie, K.M.; Philpot, R.M.; Halpert, J.R.
Orlando, Fla. : Academic Press; 1991 Nov15.
Archives of biochemistry and biophysics v. 291 (1): p.
176-186; 1991 Nov15. Includes references.
Language: English
Descriptors: Saccharomyces cerevisiae; Plasmids; Genetic transformation; Cytochromes; Gene expression; Mammals
Abstract: Three mammalian cytochromes P450 from the IIB subfamily, P450IIB11 from canine and P450IIB4 and P450IIB5 from rabbit, have been expressed in the yeast Saccharomyces cerevisiae by use of an autonomously replicating vector containing the galactose-inducible gal10 promoter. Cytochromes P450IIB4 and P450IIB5 are closely related proteins, with only 11 amino acid substitutions between them. P450IIB11 is a homologous protein, likely orthologous with IIB4 or IIB5, with 102 amino acid substitutions compared with the P450IIB4 protein and 106 compared with the P450IIB5 protein. The expressed proteins are functional in yeast microsomes, exhibiting activity toward androstenedione, 7-ethoxycoumarin, and, in some cases, progesterone. Expressed cytochromes P450IIB4 and P450IIB11 hydroxylate androstenedione with regioand stereoselectivity characteristic of the purified, reconstituted proteins. A striking difference in the androstenedione metabolite profiles of IIB4 and IIB5 was observed, with IIB4 producing almost exclusively the 16 betahydroxy metabolite and IIB5 producing the 16 alpha-hydroxy and 15 alpha-hydroxy products. This is the first time that 15 alpha-hydroxylase activity has been associated with IIB4/IIB5. This activity has also been detected in liver microsomes from some, but not all, individual phenobarbital-induced rabbits tested and is largely inhibited by anti-rabbit P450IIB immunoglobulin G. These studies illustrate the utility of the yeast expression system for defining catalytic activities of individual mammalian cytochromes P450 and identifying new marker activities that can be utilized in liver microsomes.
69 NAL Call. No.: 381 J824
Functional expression of plant plasma membrane H+-ATPase in
yeast endoplasmic reticulum.
Villalba, J.M.; Palmgren, M.G.; Berberian, G.E.; Ferguson, C.;
Serrano, R. Baltimore, Md. : American Society for Biochemistry
and Molecular Biology; 1992 Jun15.
The Journal of biological chemistry v. 267 (17): p.
12341-12349; 1992 Jun15. Includes references.
Language: English
Descriptors: Arabidopsis thaliana; Plasma membranes; Recombinant DNA; Adenosinetriphosphatase; Plasmids; Yeasts; Transformation
Abstract: Recombinant plant plasma membrane H+-ATPase has been produced in a yeast expression system comprising a multicopy plasmid and the strong promoter of the yeast PMA1 gene. Western blotting with a specific monoclonal antibody showed that the plant ATPase is one of the major membrane proteins made by the transformed cells, accounting for about 1% of total yeast protein. The plant ATPase synthesized in yeast is fully active. It hydrolyzes ATP, pumps protons, and the reaction cycle involves a phosphorylated intermediate. Phosphorylation is possible from both ATP and Pi. Unlike the situation in plants, however, most of the plant ATPase is not expressed in the yeast plasma membrane. Rather, the enzyme appears to remain trapped at a very early stage of secretory pathway: insertion into the endoplasmic reticulum. This organelle was observed to proliferate in the form of stacked membranes surrounding the yeast nucleus in order to accommodate the large amount of plant ATPase produced. In this location, the plant ATPase can be purified with high yield (70 mg from 1 kg of yeast) from membranes devoid of endogenous yeast plasma membrane H+-ATPase. This convenient expression system could be useful for other eukaryotic membrane proteins and ATPases.
70 NAL Call. No.: SB732.6.M65
The Fusarium solani-induced expression of a pea gene family
encoding high cysteine content proteins.
Chiang, C.C.; Hadwiger, L.A.
St. Paul, Minn. : APS Press; 1991 Jul.
Molecular plant-microbe interactions : MPMI v. 4 (4): p.
324-331; 1991 Jul. Includes references.
Language: English
Descriptors: Pisum sativum; Fusarium solani f.sp. phaseoli; Fusarium solani f.sp. pisi; Host parasite relationships; Pathogenesis-related proteins; Plant proteins; Cysteine; Molecular weight; Stress response; Gene expression; Genes; Disease resistance; Genetic analysis; Messenger RNA; Dna; Amino acid sequences; Nucleotide sequences
71 NAL Call. No.: 448.8 C162
Gene expression associated with light-induced conidiation in
Colletotrichum graminicola.
Yang, Z.F.; Panaccione, D.G.; Hanau, R.M.
Ottawa : National Research Council of Canada; 1991 Feb.
Canadian journal of microbiology v. 37 (2): p. 165-167. ill;
1991 Feb. Includes references.
Language: English
Descriptors: Zea mays; Colletotrichum graminicola; Conidia; Asexual reproduction; Gene expression
72 NAL Call. No.: 381 J824
Half-calmodulin is sufficient for cell proliferation.
Expressions of N-and C-terminal halves of calmodulin in the
yeast Saccharomyces cerevisiae. Sun, G.H.; Ohya, Y.; Anraku,
Y.
Baltimore, Md. : American Society for Biochemistry and
Molecular Biology; 1991 Apr15.
The Journal of biological chemistry v. 266 (11): p. 7008-7015;
1991 Apr15. Includes references.
Language: English
Descriptors: Saccharomyces cerevisiae; Calmodulin; Gene expression; Cell division; Structure activity relationships
Abstract: Calmodulin (CaM) has been shown to be an essential component for progression of nuclear division in the yeast Saccharomyces cerevisiae (Ohya, Y., and Anraku, Y. (1989) Curr. Genet. 15, 113-120). To define the functional domain of the molecule required for cell proliferation, we constructed plasmids expressing a series of N- and C-terminal halves of the CaM under the control of the galactose-inducible GAL1 promoter. These plasmids were introduced into a cmd1-disrupted yeast haploid strain, and the growth properties of the cells depending on the half-CaMs were examined. Plasmids expressing the N-terminal half (Ser1-Leu176) and the C-terminal half (Leu85-Cys147), which each maintain two complete EF-hand structures, complemented the growth defect of the cmd1 null mutation, whereas those expressing shorter regions of C- and N-terminal CaM did not. The half-CaMs that complemented the cmd1 null mutation were found to be approximately 6-fold overexpressed relative to expression of native Cam by the wild-type CMD1 gene. The levels of expression of the half CaMs with the true CMD1 promoter were not sufficient for complementation. These results demonstrate that half-CaMs (either the N- or the C-terminal) are capable of supporting growth of yeast cells when they are suitably overproduced. Cells depending solely on half-cams all showed a temperaturesensitive growth phenotype, suggesting that half-CaMs cannot carry out all the cellular functions of the complete CaM molecule.
73 NAL Call. No.: 381 AR2 Heterologous expression and characterization of an active lignin peroxidase from Phanerochaete chrysosporium using recombinant baculovirus. Johnson, T.M.; Li, J.K.K. Orlando, Fla. : Academic Press; 1991 Dec. Archives of biochemistry and biophysics v. 291 (2): p. 371-378; 1991 Dec. Includes references.
Language: English
Descriptors: Phanerochaete chrysosporium; Lignin; Ligninolytic microorganisms; Peroxidases; Isoenzymes; Genetic engineering
Abstract: The cDNA clone lambdaML-1 encoding one of the extracellular lignin peroxidases from the white rot fungus, Phanerochaete chrysosporium, was heterologously expressed in an active form using a recombinant baculovirus system. The glycosylated extracellular form of the recombinant protein contained the ferriprotoporphyrin IX moiety and was capable of oxidizing both iodide and the model lignin compound, veratryl alcohol. In comparative peroxidase assays using guaiacol and MN(II), the recombinant lignin peroxidase did not appear to be MN(II) dependent. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis demonstrated that the heterologously expressed peroxidase had an apparent molecular weight similar to that of the native fungal isozyme H8. The elution profile of the active recombinant enzyme derived by ion-exchange chromatography and immunoblot analysis using an anti-H8 monoclonal antibody provided further evidence that the lambdaML-1 DNA encodes the lignin peroxidase H8.
74 NAL Call. No.: 442.8 B5236
Heterologous expression of active manganese peroxidase from
Phanerochaete chrysosporium using the baculovirus expression
system.
Pease, E.A.; Aust, S.D.; Tien, M.
Orlando, Fla. : Academic Press; 1991 Sep16.
Biochemical and biophysical research communications v. 179
(2): p. 897-903; 1991 Sep16. Includes references.
Language: English
Descriptors: Phanerochaete chrysosporium; Manganese; Oxidoreductases; Isoenzymes; Autographa californica; Nuclear polyhedrosis viruses; Recombination; Enzyme activity; Gene expression; Purification; Characterization
Abstract: The cDNA encoding Mn peroxidase isozyme H4 from Phanerochaete chrysosporium was recombined into a baculovirus and heterologously expressed in Sf9 cells. The recombinant Mn peroxidase has the same molecular weight as the native enzyme as determined by SDS-PAGE and cross-reacts with a Mn peroxidase-specific antibody. The recombinant enzyme has a slightly lower pI than the native fungal isozyme H4 indicating some differences in post-translational modification. Phenol red, guaiacol, and vanillylacetone, substrates of the native Mn peroxidase, are oxidized by the recombinant enzyme. All of the activities axe dependent on both Mn (II) and H2O2.
75 NAL Call. No.: 381 J824
Heterologous expression of peptide hormone precursors in the
yeast Saccharomyces cerevisiae. Evidence for a novel
prohormone endoprotease with specificity for monobasic amino
acids.
Bourbonnais, Y.; Danoff, A.; Thomas, D.Y.; Shields, D.
Baltimore, Md. : American Society for Biochemistry and
Molecular Biology; 1991 Jul15.
The Journal of biological chemistry v. 266 (20): p.
13203-13209; 1991 Jul15. Includes references.
Language: English
Descriptors: Saccharomyces cerevisiae; Somatostatin; Gene expression; Plasmids; Cloning; Precursors; Mammals
Abstract: The peptide somatostatin (SRIF) exists as two different molecular species. In addition to the most common form, which is a 14-residue peptide, there is also a 14-amino acid amino-terminally extended form of the tetradecapeptide, SRIF-28. Both peptides are synthesized as larger precursors containing paired basic and monobasic amino acids at their processing sites, which, upon cleavage, generate either SRIF-14 or -28, respectively. In mammals a single prepro-SRIF molecule undergoes tissue-specific processing to generate the mature hormone whereas in some species of fish separate genes encode two distinct but homologous precursors prepro-SRIF-I and -II that give rise to SRIF-14 and -28, respectively. To investigate the molecular basis for differential processing of the prohormones we introduce their cDNAs into yeast cells (Saccharomyces cerevisiae). The signal peptides of both precursors were poorly recognized by the yeast endoplasmic reticulum translocation apparatus, consequently only low levels of SRIF peptides were synthesized. To circumvent this problem a chimeric precursor consisting of the alpha-factor signal peptide plus 30 residues of the proregion was fused to pro-SRIF-II. This fusion protein was efficiently transported through the yeast secretory pathway and processed to SRIF-28 exclusively, which is identical to the processing of the native precursor in pancreatic islet D-cells. Most significantly, cleavage of the precursor to SRIF-28 was independent of the Kex 2 endoprotease since processing occurred efficiently in a kex 2 mutant strain. We conclude that in addition to the Kex 2 protease, yeast possess a distinct prohormone converting enzyme with specificity toward monobasic processing sites.
76 NAL Call. No.: Q320.B56
Heterologous gene expression in methylotrophic yeast.
Tschopp, J.F.; Cregg, J.M.
Stoneham, Mass. : Butterworth Publishers; 1991.
Biotechnology (18): p. 305-322; 1991. In the series analytic:
Biology of Methylotrophs / edited by Israel Goldberg and J.
Stephen Rokem. Includes references.
Language: English
Descriptors: Yeasts; Substrates; Methanol; Gene expression; Genetic transformation; Fermentation
77 NAL Call. No.: 442.8 AR26
High level expression of isocitrate lyase gene of n-alkaneutilizing
yeast Candida tropicalis in Saccharomyces
cerevisiae.
Oda, K.; Atomi, H.; Ueda, M.; Kondo, J.; Teranishi, Y.;
Tanaka, A. Berlin, W. Ger. : Springer International; 1991.
Archives of microbiology v. 156 (6): p. 439-443; 1991.
Includes references.
Language: English
Descriptors: Candida tropicalis; Saccharomyces cerevisiae; Alkanes; Organelles; Isocitrate lyase; Microbial proteins; Promoters; Recombination; Genetic code; Gene expression; Enzyme activity
Abstract: The genomic DNA of peroxisomal isocitrate lyase (ICL) isolated from an n-alkane-assimilating yeast, Candida tropicalis, was truncated to utilize the original open reading frame under the control of the GAL7 promoter and was expressed in Saccharomyces cerevisiae. The recombinant ICL was synthesized as a functionally active enzyme with a specific activity, similar to the enzyme purified from C. tropicalis, and was accounted for approximately 30% of the total extractable proteins in the yeast cells. This recombinant enzyme was easily purified to homogeneity. N-terminal amino acid sequence, molecular masses of native form and subunit. amino acid composition, peptide maps, and kinetic parameters of the recombinant ICL were essentially the same its those of ICL purified from C. tropicalis. From these facts, S. cerevisiae was suggested to be an excellent microorganism to highly express the genes encoding peroxisomal proteins of C. tropicalis.
78 NAL Call. No.: QH426.C8 The identification of 18 nuclear genes required for the expression of the yeast mitochondrial gene encoding cytochrome c oxidase subunit 1. Pel, H.J.; Tzagoloff, A.; Grivell, L.A. Berlin, W. Ger. : Springer International; 1992. Current genetics v. 21 (2): p. 139-146; 1992. Includes references.
Language: English
Descriptors: Saccharomyces cerevisiae; Mitochondrial DNA; Genes; Cytochrome-c oxidase; Gene expression; Genetic regulation; Nuclei; Nucleocytoplasmic interaction; Induced mutations; Mutants; Messenger RNA; Rna editing; Transcription; Mitochondrial genetics
Abstract: Eighteen nuclear mutants of the yeast Saccharomyces cerevisiae, each disturbed in the biosynthesis of the mitochondrially encoded cytochrome c oxidase subunit 1 (cox1) and each representing a distinct complementation group, have been examined to identify the level at which COX1 expression is affected. RNA blotting revealed that most have a defect in the processing of COX1 precursor-mRNA; only a few are defective in COX1 transcription and/or pre-mRNA stability. In most RNA-processing mutants, the absence of the COX1 messenger results from a defect in the splicing of one or more COX1 introns. In turn, this defect can be ascribed to a mutation in a nuclear gene which is either directly involved in splicing or else acts indirectly by impairing COX1 translation.
79 NAL Call. No.: 500 N21P
Identification of a tomato gene for the ethylene-forming
enzyme by expression in yeast.
Hamilton, A.J.; Bouzayen, M.; Grierson, D.
Washington, D.C. : The Academy; 1991 Aug15.
Proceedings of the National Academy of Sciences of the United
States of America v. 88 (16): p. 7434-7437; 1991 Aug15.
Includes references.
Language: English
Descriptors: Lycopersicon esculentum; Saccharomyces cerevisiae; Amino acid sequences; Biosynthesis; Ethylene; Enzyme activity; Gene expression; Genetic transformation; Molecular genetics; Nucleotide sequences; Oxidoreductases
Abstract: The ethylene-forming enzyme (EFE), which catalyzes the last step in the biosynthesis of the plant hormone ethylene, has never been purified and no molecular probes are available. Recently, a putative cDNA done for tomato EFE (pTOM13) has been identified by inhibiting ethylene synthesis with an antisense gene expressed in transgenic plants. A direct test of its function has been made by expression of a pTOM13 gene in Saccharomyces cerevisiae. After cloning artefacts were discovered in the 5' region of the cDNA, a corrected cDNA (pRC13) was created by the fusion of the 5' end of a genomic clone to the 3' end of the cDNA and expressed in S. cerevisiae. Cultures of transformed yeast converted 1- aminocyclopropane-l-carboxylic acid (ACC) to ethylene, whereas control cells did not. This EFE activity displays similar characteristics to EFE found in plant tissue: it converts the trans isomer of the ACC analogue 1-amino-2-ethylcyclopropanel -carboxylic acid to 1-butene in preference to the cis isomer, and it is strongly inhibited by cobaltous ions and 1,10- phenanthroline. Furthermore, information gained from the activity of effectors on yeast EFE activity supports the hypothesis that EFE is one of a group of hydroxylase enzymes.
80 NAL Call. No.: 500 N21P
Identification of Dictyostelium Galpha genes expressed during
multicellular development.
Hadwiger, J.A.; Wilkie, T.M.; Strathmann, M.; Firtel, R.A.
Washington, D.C. : The Academy; 1991 Sep15.
Proceedings of the National Academy of Sciences of the United
States of America v. 88 (18): p. 8213-8217; 1991 Sep15.
Includes references.
Language: English
Descriptors: Dictyostelium; Amino acid sequences; Binding proteins; Gene expression; Gene mapping; Genetic analysis; Genetic code; Guanine; Nucleotide sequences; Polymerase chain reaction
Abstract: Guanine nucleotide-binding protein (G protein)- mediated signal transduction constitutes a common mechanism by which cells receive and respond to a diverse set of environmental signals. Many of the signals involved in the developmental life cycle of the slime mold Dictyostelium have been postulated to be transduced by such pathways and, in some cases, these pathways have been demonstrated to be dependent on specific G proteins. Using the polymerase chain reaction, we have identified two additional Dictyostelium G alpha genes, G alpha 4 and G alpha 5, that are developmentally regulated. Transcripts from both of these genes are primarily expressed during the multicellular stages of development, suggesting possible roles in cell differentiation or morphogenesis. The entire G alpha 4 gene was sequenced and found to encode a protein consisting of 345 amino acids. The G alpha 4 subunit is homologous to other previously identified G alpha subunits, including the Dictyostelium G alpha 1 (43% identity) and G alpha 2 (41% identity) subunits. However, the G alpha 4 subunit contains some unusual sequence divergences in residues highly conserved among most eukaryotic G alpha subunits, suggesting that G alpha 4 may be a member of another class of G alpha subunits.
81 NAL Call. No.: QH442.A1G4 Improved shuttle vectors for cloning and high-level Cu2+- mediated expression of foreign genes in yeast. Macreadie, I.G.; Horaitis, O.; Verkuylen, A.J.; Savin, K.W. Amsterdam : Elsevier Science Publishers; 1991. Gene v. 104 (1): p. 107-111; 1991. Includes references.
Language: English
Descriptors: Saccharomyces cerevisiae; Recombinant DNA; Gene expression; Vectors
Abstract: New yeast episomal vectors having a high degree of utility for cloning and expression in Saccharomyces cerevisiae are described. One vector, pYEULlacZ, is based on pUC19 and employs the pUC19 multiple cloning site for the selection of recombinants in Escherichia coli by lacZ inactivation. In addition, the vector contains two genes, URA3 and leu2-d, for selection of the plasmid in ura3 or leu2 yeast strains. The presence of the leu2-d gene appears to promote replication at high copy numbers. The introduction of CUP1 cassettes allows these plasmids to direct Cu2+-regulated production of foreign proteins in yeast. We show the production of a helminth antigen as an example of the vector application.
82 NAL Call. No.: 500 N21P In vitro assembly of apophytochrome and apophytochrome deletion mutants expressed in yeast with phycocyanobilin. Deforce, L.; Tomizawa, K.I.; Ito, N.; Farrens, D.; Song, P.S.; Furuya, M. Washington, D.C. : The Academy; 1991 Dec01. Proceedings of the National Academy of Sciences of the United States of America v. 88 (23): p. 10392-10396. ill; 1991 Dec01. Includes references.
Language: English
Descriptors: Saccharomyces cerevisiae; Yeasts; Mutants; Recombination; Gene expression; Phytochrome; Biosynthesis; Amino acid sequences
Abstract: Recombinant pea type I phytochrome apoprotein expressed in yeast is shown to assemble in vitro with phycocyanobilin to produce a photoreversible phytochrome-like adduct. As an initial investigation of the amino acid sequence requirements for chromophore incorporation, three phyA gene product deletion mutants were produced in yeast. Truncation of the N-terminal tail to residue 46 demonstrates that this region is not critical to bilin attachment, but a deletion mutant lacking 222 amino acids from the N terminus failed to yield holophytochrome in vitro, under the same conditions. A mutant comprising a deletion of the C terminus to residue 548 showed bilin incorporation and red/far-red photoreversibility, indicating that bilin-apophytochrome assembly still occurred even when the entire C-terminal domain was truncated.
83 NAL Call. No.: 442.8 Z34
Inhibition of cell proliferation in Saccharomyces cerevisiae
by expression of human NAD+ ADP-ribosyltransferase requires
the DNA binding domain ("zinc fingers").
Kaiser, P.; Auer, B.; Schweiger, M.
Berlin, W. Ger. : Springer International; 1992 Mar.
M G G : Molecular and general genetics v. 232 (2): p. 231-239;
1992 Mar. Includes references.
Language: English
Descriptors: Saccharomyces cerevisiae; Man; Gene transfer; Genetic transformation; Pentosyltransferases; Genes; Gene expression; Dna; Binding site; Cell division; Chemical reactions; Adp; Ribose; Enzyme activity; Nucleoproteins
Abstract: Constitutive expression of human nuclear NAD+: protein ADP-ribosyltransferase (polymerizing) [pADPRT; poly(ADP-ribose)polymerase; EC 2.4.2.30] as an active enzyme in Saccharomyces cerevisiae, under the control of the alcohol dehydrogenase promoter, was only possible with simultaneous inhibition of ADP-ribosylation by 3-methoxybenzamide. Induction of fully active pADPRT from the inducible galactose epimerase promoter resulted in inhibition of cell division and morphological changes reminiscent of cell cycle mutants. Expression of a pADPRT cDNA truncated at its 5' end had no influence on cell proliferation at all. Obviously the aminoterminal part of the DNA binding domain containing the first "zinc finger", which is essential for inducibility of pADPRT activity by DNA breaks, is also required for inhibition of cell growth on expression in yeast. Full-length as well as truncated pADPRT molecules were directed to the cell nucleus where the fully active enzyme produced large amounts of poly(ADP-ribose) by automodification. Since pADPRT turned out to be the only target for ADP-ribosylation in these cells, elevated levels of poly(ADP-ribose) were the most likely cause of inhibition of cell division, presumably resulting from interaction with chromosomal proteins.
84 NAL Call. No.: QH442.A1G4
The K2-type killer toxin- and immunity-encoding region from
Saccharomyces cerevisiae: structure and expression in yeast.
Meskauskas, A.; Citavicius, D.
Amsterdam : Elsevier Science Publishers; 1992.
Gene v. 111 (1): p. 135-139; 1992. Includes references.
Language: English
Descriptors: Saccharomyces cerevisiae; Rna; Mycotoxins; Phenotypes; Cloning; Nucleotide sequences; Amino acid sequences; Plant proteins; Genetic code; Resistance
Abstract: The cDNA copies of M2-1, the larger heat-cleavage product of M2 double-stranded (ds) RNA, have been synthesized, cloned, sequenced and expressed in yeast. This sequence, in combination with the known terminal sequence of M2-1 dsRNA, identifies a translation reading frame for a 362-amino-acid protein of 38.7 kDa, similar in size to the one of several protein species produced from M2-1 dsRNA in vitro translation. The expression of this cDNA clone in yeast confers both killer and immunity phenotypes.
85 NAL Call. No.: 448.3 AP5
Manganese regulation of manganese peroxidase expression and
lignin degradation by the white rot fungus Dichomitus
squalens.
Perie, F.H.; Gold, M.H.
Washington, D.C. : American Society for Microbiology; 1991
Aug. Applied and environmental microbiology v. 57 (8): p.
2240-2245; 1991 Aug. Includes references.
Language: English
Descriptors: Polyporus; Lignin; Microbial degradation; Manganese; Peroxidases; Laccase; Enzyme activity; Oxidoreductases; Mineralization
Abstract: Extracellular manganese peroxidase and laccase activities were detected in cultures of Dichomitus squalens (Polyporus anceps) under conditions favoring lignin degradation. In contrast, neither extracellular lignin peroxidase nor aryl alcohol oxidase activity was detected in cultures grown under a wide variety of conditions. The mineralization of 14C-ring-, -side chain-, and -methoxylabeled synthetic guaiacyl lignins by D. squalens and the expression of extracellular manganese peroxidase were dependent on the presence of Mn(II), suggesting that manganese peroxidase is an important component or this organism's lignin degradation system. The expression of laccase activity was independent of manganese. In contrast to previous findings with Phanerochaete chrysosporium, lignin degradation by D. squalens proceeded in the cultures containing excess carbon and nitrogen.
86 NAL Call. No.: QH426.C8
The mating type in fission yeast is switched independently of
its expression. Ruusala, T.
Berlin, W. Ger. : Springer International; 1991.
Current genetics v. 20 (5): p. 379-383; 1991. Includes
references.
Language: English
Descriptors: Endomycetales; Loci; Mating; Genetic change; Gene expression; Transcription; Transposable elements; Plasmids
Abstract: The mating type of fission yeast is determined by the mat1 locus on chromosome II. The sequence content of this locus, and hence the mating type, is switched in a strictly regular pattern by transposition from one of two unexpressed mating type sequences. The expressed and the two silent sequences are located on the same chromosome. It is not understood how one of the two donor sequences is selected in this reaction. Here I test the possibility that the selection is governed by gene expression from the mat1 locus. Such a mechanism could favor transposition of a donor sequence of opposite mating type to the one present at mat1. Alternatively it could disfavor transposition of a synonymous sequence. The present data argue strongly against any type of participation of mat1 gene products in the choice of donor during the mating type switch. Alternative steering mechanisms are discussed.
87 NAL Call. No.: QR53.B56
Methanol as a convenient inducer of heat shock elementdirected
heterologous gene expression in yeast.
Kirk, N.; Piper, P.W.
Middlesex : Science and Technology Letters; 1991 Jul.
Biotechnology letters v. 13 (7): p. 465-470; 1991 Jul.
Includes references.
Language: English
Descriptors: Saccharomyces cerevisiae; Heat shock; Methanol; Induction; Tolerance; Survival; Promoters; Reporter genes; Genetic transformation; Gene expression; Fermentation
Abstract: The addition of sublethal concentrations of methanol to yeast cultures was found to give strong induction of a lacZ gene under heat shock element (HSE) control. Methanol addition is therefore an easily-executed alternative to temperature upshift for the induction of HSE-directed gene expression, although in this study it gave slightly lower levels of induction compared to heat shock.
88 NAL Call. No.: QK600.B72
Methylotrophic yeasts as gene expression systems.
Veale, R.A.; Sudberry, P.E.
Cambridge : Cambridge University Press; 1991.
Symposium series - British Mycological Society (18): p.
118-128; 1991. In the series analytic: Applied molecular
genetics of fungi / edited by J. F. Peberdy, C. E. Caten, J.
E. Ogden and J. W. Bennett. Symposium of the British
Mycological Society held at the University of Nottingham,
April 1990. Literature review. Includes references.
Language: English
Descriptors: Pichia; Hansenula; Genetic transformation; Gene transfer; Gene expression; Structural genes; Proteins; Plasmids; Vectors; Alcohol oxidoreductases; Promoters; Methanol; Substrates; Literature reviews
89 NAL Call. No.: QP501.B64
Mitochondrial gene expression in Saccharomyces cerevisiae:
proteolysis of nascent chains in isolated yeast mitochondria
optimized for protein synthesis. Black-Schaefer, L.; McCourt,
J.D.; Poyton, R.O.; McKee, E.E. London : The Biochemical
Society; 1991 Feb15.
The Biochemical journal v. 274 (pt.1): p. 199-205; 1991 Feb15.
Third in a series. Includes references.
Language: English
Descriptors: Saccharomyces cerevisiae; Mitochondria; Protein synthesis; Translation; Proteolysis; Gene expression
Abstract: We demonstrate here that mitochondrial translation products synthesized by isolated yeast mitochondria are subject to rapid proteolysis. The loss of label from mitochondrial peptides synthesized in vitro comes from two distinct pools of peptides: one that is rapidly degraded (t1/2 of minutes) and one that is much more resistant to proteolysis (t1/2 of hours). As the length of the incubation period increases, the percentage of labelled peptides in the rapidlyturning -over pool decreases and cannot be detected after 60 min of incubation. This proteolysis is inhibited by chloramphenicol and is dependent on the presence of ATP. The loss of label during the chase occurs from fully completed translation products. The proteolysis observed here markedly affects measurements of rates of mitochondrial protein synthesis in isolated yeast mitochondria. In earlier work, in which proteolysis was not considered, mitochondrial translation was thought to stop after 20-30 min of incubation. In the present study, by taking proteolysis into account, we demonstrate that the rate of translation in isolated mitochondria is actually constant for nearly 60 min and then decreases to near zero by 80 min of incorporation. These findings have allowed us to devise a procedure for measuring the 'true' rate of translation in isolated mitochondria. In addition, they suggest that mitochondrial translation products which normally assemble with nuclear-encoded gene products into multimeric enzyme complexes are unstable without their nuclear-encoded counterparts.
90 NAL Call. No.: QK1.D5 Bd.166
Mitochondriale Genexpression bei Pilzen molekulare Analysen
zur nukleo-zytoplasmatischen Wechselwirkung [Mitochondrial
gene expression in fungi].
Heinen, Ute
Berlin : J. Cramer,; 1991.
107 p. : ill. ; 23 cm. (Dissertationes Botanicae ; Bd. 166).
Originally presented as the author's thesis (doctoral).
Includes bibliographical references (p. 88-107).
Language: German
91 NAL Call. No.: SB732.6.M65
Molecular analysis of the laccase gene from the chestnut
blight fungus and selective suppression of its expression in
an isogenic hypovirulent strain. Choi, G.H.; Larson, T.G.;
Nuss, D.L.
St. Paul, Minn. : APS Press; 1992 Mar.
Molecular plant-microbe interactions : MPMI v. 5 (2): p.
119-128; 1992 Mar. Includes references.
Language: English
Descriptors: Castanea; Cryphonectria parasitica; Plant pathogenic fungi; Nucleotide sequences; Amino acid sequences; Comparisons; Neurospora crassa; Laccase; Genes; Species differences; Strain differences; Hypovirulence; Strains; Messenger RNA; Tubulin; Actin; Glyceraldehyde-3-phosphate dehydrogenase; Gene expression; Genetic analysis; Introns
92 NAL Call. No.: QK600.S4 The molecular biology of Trichoderma and its application to the expression of both homologous and heterologous genes. Nevalainen, K.M.H.; Penttila, M.E.; Harkki, A.; Teeri, T.T.; Knowles, J. New York, N.Y. : Marcel Dekker; 1991. Mycology series v. 8: p. 129-148; 1991. In the series analytic: Molecular industrial mycology: systems and applications for filamentous fungi / edited by S. A. Leong and R. M. Berka. Literature review. Includes references.
Language: English
Descriptors: Trichoderma; Molecular biology; Gene expression; Mutants; Cellulolytic microorganisms; Cellulose digestion; Cellulose; Biosynthesis; Genetic regulation; Genetic code; Literature reviews
93 NAL Call. No.: 500 N21P
Molecular cloning and expression in photosynthetic bacteria of
a soybean cDNA coding for phytoene desaturase, an enzyme of
the carotenoid biosynthesis pathway.
Bartley, G.E.; Viitanen, P.V.; Pecker, I.; Chamovitz, D.;
Hirschberg, J.; Scolnik, P.A.
Washington, D.C. : The Academy; 1991 Aug01.
Proceedings of the National Academy of Sciences of the United
States of America v. 88 (15): p. 6532-6539; 1991 Aug01.
Includes references.
Language: English
Descriptors: Glycine max; Glycine soja; Amino acid sequences; Carotenoids; Cloning; Dna; Gene expression; Gene mapping; Nucleotide sequences; Restriction fragment length polymorphism
Abstract: Carotendoids are orange, yellow, or red photoprotective pigments present in all plastids. The first carotenoid of the pathway is phytoene, a colorless compound that is converted into colored carotenoids through a series of desaturation reactions. Genes coding for carotenoid desaturases have been cloned from microbes but not from plants. We report the cloning of a cDNA for pds1, a soybean (Glycine max) gene that, based on a complementation assay using the photosynthetic bacterium Rhodobacter capsulatus, codes for an enzymic that catalyzes the two desaturation reactions that convert phytoene into zeta-carotene, a yellow, carotenoid. The 2281-base-pair cDNA clone analyzed contains an open reading frame with the capacity to code for a 572-residue protein of predicted Mr 63,851. Alignment of the deduced Pds1 peptide sequence with the sequences of fungal and bacterial carotenoid desaturases revealed conservation of several amino acid residues, including a dinucleotide-binding motif that could mediate binding to FAD. The Pds1 protein is synthesized in vitro as a precursor that, upon import into isolated chloroplasts, is processed to a smaller mature form. Hybridization of the pds1 cDNA to genomic blots indicated that this gene is a member of a low-copy-number gene family. One of them loci was genetically mapped using restriction fragment length polymorphisms between Glycine max and Glycine soja. We conclude that pds1 is a nuclear gene encoding a phytoene desaturase enzyme that, as its microbial counterparts, contains sequence motifs characteristic of flavoproteins.
94 NAL Call. No.: 500 N21P
Molecular cloning, expression, and induction of berberine
bridge enzyme, an enzyme essential to the formation of
benzophenanthridine alkaloids in the response of plants to
pathogenic attack.
Dittrich, H.; Kutchan, T.M.
Washington, D.C. : The Academy; 1991 Nov15.
Proceedings of the National Academy of Sciences of the United
States of America v. 88 (22): p. 9969-9973; 1991 Nov15.
Includes references.
Language: English
Descriptors: Eschscholzia californica; Cloning; Gene expression; Molecular genetics; Oxidoreductases; Phytoalexins; Biosynthesis; Enzyme activity; Nucleotide sequences; Saccharomyces cerevisiae; Amino acid sequences; Fungal diseases
Abstract: The berberine bridge enzyme [(S)-reticuline: oxygen oxidoreductase (methylene-bridge-forming), EC 1.5.3.9] is a vesicular plant enzyme that catalyzes the formation of the berberine bridgehead carbon of (S)-scoulerine from the Nmethyl carbon of (S)-reticuline in a specific, unparalleled reaction along the biosynthetic pathway that leads to benzophenanthridine alkaloids. Cytotoxic benzophenanthridine alkaloids are accumulated in certain species of Papaveraceae and Fumariaceae in response to pathogenic attack and, therefore, function as phytoalexins. The berberine bridge enzyme has been purified to homogeneity from elicited cellsuspension cultures of Eschscholtzia californica, and partial amino acid sequences have been determined. A cDNA, isolated from a lambda-gt11 cDNA bank of elicited E. californica cellsuspension cultures, coded for an open reading frame of 538 amino acids. The first 22 amino acids constitute the putative signal peptide. The mature protein has a Mr of 57,352, excluding carbohydrate. The berberine bridge enzyme was heterologously expressed in a catalytically active form in Saccharomyces cerevisiae. Southern hybridization with genomic DNA suggests that there is only one gene for the enzyme in the E. californica genome. Hybridized RNA blots from elicited E. californica cell-suspension cultures revealed a rapid and transient increase in poly(A)+ RNA levels that preceded both the increase in enzyme activity and the accumulation of benzophenanthridine alkaloids, emphasizing the integral role of the berberine bridge enzyme in the plant response to pathogens.
95 NAL Call. No.: QR1.F44
Molecular cloning of genes from the rumen anaerobic fungus
Neocallimastix frontalis: expression during hydrolase
induction.
Reymond, P.; Durand, R.; Hebraud, M.; Fevre, M.
Amsterdam : Elsevier Science Publishers; 1991 Jan01.
FEMS microbiology letters - Federation of European
Microbiological Societies v. 77 (1): p. 107-112; 1991 Jan01.
Includes references.
Language: English
Descriptors: Neocallimastix; Rumen fungi; Anaerobes; Hydrolases; Induction; Dna hybridization; Rna; Cloning; Gene expression; Screening; Dna libraries; Translation; Transcription; Nucleotide sequences
Abstract: Glycoside and polysaccharide hydrolase production by the rumen anaerobic fungus, Neocallimastix frontalis is induced by the presence of crystalline cellulose. A differential screening of a cDNA library was used to isolate DNA sequences transcribed at high levels under growth conditions which induce enzyme production. Seven clones were isolated that prefentially hybridized to the induced cDNA probe versus the non-induced cDNA probe. Southern analysis showed that a cDNA clone (118) hybridized to a DNA probe encoding part of the exo-cellobiohydrolase I (CBH I) gene of Trichoderma reesei. Northern analysis demonstrated that the cDNA 118 was transcribed to yield a 2.1 kb RNA. This transcript was induced in the presence of cellulose.
96 NAL Call. No.: 500 N21P Monitoring of intracellular calcium in Saccharomyces cerevisiae with an apoaequorin cDNA expression system. Nakajima-Shimada, J.; Iida, H.; Tsuji, F.I.; Anraku, Y. Washington, D.C. : The Academy; 1991 Aug01. Proceedings of the National Academy of Sciences of the United States of America v. 88 (15): p. 6878-6882; 1991 Aug01. Includes references.
Language: English
Descriptors: Saccharomyces cerevisiae; Calcium ions; Cell growth; Gene expression; Luminescence; Plant proteins; Recombinant DNA; Sex pheromones
Abstract: A method is described for measuring cytosolic free Ca2+ and its time-dependent changes in the yeast Saccharomyces cerevisiae by using the luminescent protein aequorin as a Ca2+-specific indicator. This method with intact yeast cells is labeled "in vivo" to distinguish it from methods with cell extracts, labeled "in vitro." A plasmid in which the apoaequorin cDNA was joined downstream from the glyceraldehyde-3-phosphate dehydrogenase gene promoter was constructed and introduced into yeast cells. The intracellular concentration of apoaequorin expressed by the cDNA was approximately 1 micromole, which was high enough to detect the cytosolic Ca2+. Growth of the transformed cells was normal. In the in vitro method, apoaequorin in crude cell extracts was regenerated into aequorin by mixing with coelenterazine, the substrate for the luminescence reaction, whereas in the in vivo method, aequorin was regenerated by incubating intact cells with coelenterazine. Simultaneous addition of 10 mM CaCl2 and 10 micromole A23187, a Ca2+ ionophore, to coelenterazine-incorporated cells generated luminescence. Coelenterazine-incorporated cells also responded to native extracellular stimuli. A mating pheromone, alpha-factor, added to cells of mating type a or alpha, generated extracellular Ca2+-dependent luminescence specifically in a mating type cells, with maximal intensity occurring 45-50 min after addition of alpha-factor. Glucose added to glucose-starved G0/G1 cells stimulated an increase in extracellular Ca2+- dependent luminescence with maximal intensity occurring 2 min after addition. These results show the usefulness of the aequorin system in monitoring [Ca2+]i, response to extracellular stimuli in yeast cells.
97 NAL Call. No.: 381 J824
Multiple regulatory elements control expression of the gene
encoding the Saccharomyces cerevisiae cytochrome P450,
lanosterol 14 alpha-demethylase (ERG11).
Turi, T.G.; Loper, J.C.
Baltimore, Md. : American Society for Biochemistry and
Molecular Biology; 1992 Jan25.
The Journal of biological chemistry v. 267 (3): p. 2046-2056;
1992 Jan25. Includes references.
Language: English
Descriptors: Saccharomyces cerevisiae; Cytochrome p-450; Lanosterol; Enzymes; Ergosterol; Biosynthesis; Gene expression; Genetic regulation
Abstract: The major cytochrome P450 in the yeast Saccharomyces cerevisiae, lanosterol 14 alpha-demethylase (ERG11), catalyzes an essential reaction in the biosynthesis of ergosterol, the predominant sterol of yeast. Protein levels of this cytochrome P450 are known to be affected by carbon source, oxygen, and heme, as well as the growth state of the culture. We have determined that ERG11 message levels increase during growth on glucose, in the presence of heme, and during oxygen limiting growth conditions and, unexpectedly, during anaerobic growth. To determine the cis-acting regions responsible for regulation of expression of the ERG11 promoter under optimal conditions of fermentative growth, deletion analysis was performed using the Escherichia coli lacZ as a reporter gene. Two upstream activating sequences, UAS1 and UAS2, and an upstream repressor element, URS1, plus a second possible or cryptic repressor element, URS2, were identified in the ERG11 promoter. The HAP1 protein product apparently participates in activation from UAS1 but not from UAS2. Sequences resembling ERG11 UAS2 were identified in seven additional oxygen-regulated genes. Repression of ERG11, expression was dependent upon the ROX1 repressor and additional repressor(s) designated as Old (overexpression of lanosterol demethylase). These data indicate that ERG11 is a member of the hypoxic gene family which includes ANB1, COX5b, CYC7, and HEM13. Furthermore, NADPH-cytochrome P450 reductase (CPR1), another component in this P450 system, appears to be coordinately regulated with ERG11.
98 NAL Call. No.: QH442.A1G4 A
novel, highly regulated, rapidly inducible system for the
expression of chicken progesterone receptor, cPRA, in
Saccharomyces cerevisiae. Poletti, A.; Weigel, N.L.;
McDonnell, D.P.; Schrader, W.T.; O'Malley, B.W.; Conneely,
O.M.
Amsterdam : Elsevier Science Publishers; 1992.
Gene v. 114 (1): p. 51-58; 1992. Includes references.
Language: English
Descriptors: Saccharomyces cerevisiae; Genetic engineering; Promoters; Plant proteins; Recombinant DNA; Hormone receptors; Progesterone; Genes; Fowls; Gene expression; Transcription; Controlling elements
Abstract: A rapidly inducible and tightly regulated system for the expression of protein in yeast is based on a chimeric promoter constructed of two copies of a vitellogenin-estrogenresponse element (ERE) which are inserted upstream from the promoter of the yeast gene encoding iso-1-cytochrome c. The chimeric promoter was inserted in a yeast expression plasmid upstream from the coding sequence of ubiquitin fused in frame to a cDNA encoding the full-length chicken progesterone receptor A (cPR(A)). The resultant plasmid (YEpA2) was cotransformed in Saccharomyces cerevisiae with a plasmid which encodes the human estrogen receptor. Estradiol (E2)-induced transactivation of the chimeric promoter results in transcription of the cPR(A) gene from YEpA2 and synthesis of cPR(A). The fusion protein, ubiquitin-cPR(A), is rapidly cleaved in vivo to produce cPR(A). Analysis of samples by Western immunoblot shows that cPR(A) is almost undetectable in the absence of E2, and that treatment with 50 nM E2 results in a 500-1000-fold induction of cPR(A) (0.06-0.3% of the total protein) after 1 h. The plasmid-expressed soluble receptor is stable and demonstrates the correct affinity for its ligand. We have prepared yeast extracts using enzymatic digestion of the cell wall with oxalyticase followed by hypotonic shock. This has resulted in a dramatic increase in the % of receptor which binds hormone compared to previous studies which used mechanical disruption techniques. The cPR(A) is biologically active since it activates transcription of a co-transformed reporter gene containing its response element. This system is well suited for the regulated synthesis of proteins in yeast and may be especially useful for producing labile proteins or proteins toxic to the cells.
99 NAL Call. No.: QH431.A1G43
Nuclear-cytoplasmic interactions in the resistance of wheat to
fungal pathogens. II. Effects of cultivated and wild cereal
cytoplasms on the expression of the genome of the Leningradka
variety during interaction with the floury mildew pathogen.
Voluevich, E.A.; Buloichik, A.A.
New York, N.Y. : Consultants Bureau; 1992 Jun.
Soviet genetics v. 27 (12): p. 1501-1505; 1992 Jun.
Translated from: Genetika, v.27 (12), 1991, p. 2103-2108.
(QH431.A1G4). Includes references.
Language: English; Russian
Descriptors: Triticum aestivum; Aegilops squarrosa; Aegilops ovata; Triticum spelta; Plant pathogenic fungi; Plant breeding; Substitution lines; Genetic resistance; Mildews; Fungal diseases; Nucleocytoplasmic interaction; Cytoplasmic inheritance; Plant development; Crop growth stage; Sporulation
Abstract: The quantitative resistance of three alloplasmatic and 29 substituted soft wheat lines containing the nuclear genome of the Leningradka variety to a floury mildew population was studied on a stringent infective background, by measuring the mean spore-bearing ability of the fungus per cm2 of the third and flag leaves of each plant line and the initial Leningradka variety. Dispersion analysis showed that the plant cytoplasm had a significant effect on pathogen reproduction during the ontogenetic development of wheat plants. Cytoplasms from a number of cultivated and wild species had modifying effects on the expression of the nuclear genes controlling quantitative resistance. It is suggested that a specific interaction between the fungal and host plant genomes is involved in determining quantitative resistance in the presence of cytoplasms modifying fungal reproduction during plant ontogenesis. A non-specific interaction probably occurs in the presence of cytoplasms maintaining a stable level of quantitative resistance during ontogenesis.
100 NAL Call. No.: QH426.C8
Optimisation of a host/vector system for heterologous gene
expression by Hansenula polymorpha.
Sierkstra, L.N.; Verbakel, J.M.A.; Verrips, C.T.
Berlin, W. Ger. : Springer International; 1991.
Current genetics v. 19 (2): p. 81-87; 1991. Includes
references.
Language: English
Descriptors: Endomycetales; Vectors; Plasmids; Gene transfer;
Gene expression; Cyamopsis tetragonoloba; Alpha-galactosidase;
Reporter genes; Saccharomyces cerevisiae; Betafructofuranosidase;
Genetic transformation; Nucleotide
sequences; Oxidoreductases
Abstract: For the methylotrophic yeast, Hansenula polymorpha, expression vectors with different origins of replication have been constructed in order to analyse their influence on transformation and integration efficiency. The constructed plasmids are identical except for their origin of replication, which involve, respectively, that of the Saccharomyces cerevisiae 2-mu m plasmid and a H. polymorpha ARS sequence (HARS2). A plasmid with no origin of replication served as a control. The plasmids also contained the alpha-galactosidase expression cassette, consisting of the Cyamopsis tetragonoloba alpha-galactosidase gene, the H. polymorpha methanol oxidase promoter and terminator, and the S. cerevisiae invertase signal sequence. The transformation frequencies of the expression vectors containing the 2-mu m and the HARS2 origins of replication, and no origin of replication, were 2, 50 and 15 per microgram of DNA respectively, which demonstrates the negative effect of the 2-mu m sequence on the transformation frequency. Autonomously replicating plasmids could be isolated from the transformants obtained with the plasmid containing either the 2-mu m or the HARS2 sequence. Integration of the 2- mu m based plasmid into the H. polymorpha genome could not be established using a standard procedure. This is in contrast with transformants containing a plasmid bearing the HARS2 sequence or else with no origin of replication, which shows that the 2-mu m sequence negatively influences the integration of the expression vector into the H. polymorpha genome. Integration of expression plasmids occurred in 50% of the analysed integrants on the homologous methanol oxidase locus, and tandem integration was favoured. The level of specific mRNA, and the expression of the alpha-galactosidase protein by these integrants, was proportional to the number of integrated copies of the expression plasmid in the H. polymorpha genome.
101 NAL Call. No.: 442.8 Z34
The organization and expression of the Saccharomyces
cerevisiae L4 ribosomal protein genes and their identification
as the homologues of the mammalian ribosomal protein gene L7a.
Yon, J.; Giallongo, A.; Fried, M.
Berlin, W. Ger. : Springer International; 1991 May.
M G G : Molecular and general genetics v. 227 (1): p. 72-80;
1991 May. Includes references.
Language: English
Descriptors: Saccharomyces cerevisiae; Genes; Ribosomes; Proteins; Cloning; Nucleotide sequences; Amino acid sequences; Transcription; Messenger RNA; Comparisons; Mice
Abstract: A cDNA for the mouse ribosomal protein (rp) L7a, formerly called Surf-3, was used as a probe to isolate two homologous genes from Saccharomyces cerevisiae. The two yeast genes (L4-1 and L4-2) were identified as encoding S. cerevisiae L4 by 2D gel analysis of the product of the in vitro translation of hybrid-selected mRNA and additionally by direct amino acid sequencing. The DNA sequences of the two yeast genes were highly homologous (95%) over the 771 bp that encode the 256 amino acids of the coding regions but showed little homology outside the coding region. L4-1 differed from L4-2 by 7 out of the 256 amino acids in the coding region, which is the greatest divergence between the products of any two duplicated yeast ribosomal protein genes so far reported. There is strong homology between the mouse rpL7a/Surf-3 and the yeast L4 genes -57% at the nucleic acid level and also 57% at the amino acid level (though some regions reach as much as 80-90% homology). While most yeast ribosomal protein genes contain an intron in their 5' region both L4-1 and L4-2 are intronless. The mRNAs derived from each yeast gene contained heterogenous 5' and 3' ends but in each case the untranslated leaders were short. The L4-1 mRNA was found to be much more abundant than the L4-2 mRNA as assessed by cDNA and transcription analyses. Yeast cells containing a disruption of the L4-1 gene formed much smaller colonies than either wildtype or disrupted L4-2 strains. Disruption of both L4 genes is a lethal event, probably due to an inability to produce functional ribosomes.
102 NAL Call. No.: QH426.C8
Over-expression, purification and determination of the
proteolytic processing site of the yeast mitochonrial CBS1
protein.
Korte, A.; Michaelis, U.; Lottspeich, F.; Rodel, G.
Berlin, W. Ger. : Springer International; 1991.
Current genetics v. 20 (1/2): p. 87-90; 1991. Includes
references.
Language: English
Descriptors: Saccharomyces cerevisiae; Mitochondria; Plant proteins; Purification; Amino acid sequences; Proteolysis; Genes; Genetic transformation; Gene expression
Abstract: Yeast transformants harboring the CBS1 gene under the control of the strong ADC1 promoter on a high copy number plasmid express the mitochondrial CBS1 protein at artificially high levels. Over-expressed protein is imported into mitochondria and correctly processed to yield the mature mitochondrial 23.5 kDa form, but differs in its solubility properties from CBS1 in wild-type mitochondria. It forms insoluble protein aggregates, which are refractory to solubilization with 1% Taurodeoxycholate. We exploited this observation to separate CBS1 from the bulk of mitochondrial proteins and to isolate CBS1 after SDS gel electrophoresis. Determination of the amino-terminal amino acids of the purified protein reveals that the mature CBS1 protein starts with Ile(30), at the characteristic distance of +2 amino acids from an arginine residue (Arg(28)). The cleavage site shows a remarkable homology to that of subunit 9 of the F0F1 ATPase from Neurospora crassa.
103 NAL Call. No.: QH426.C8
PDC6, a weakly expressed pyruvate decarboxylase gene from
yeast, is activated when fused spontaneously under the control
of the PDC1 promoter. Hohmann, S.
Berlin, W. Ger. : Springer International; 1991.
Current genetics v. 20 (5): p. 373-378; 1991. Includes
references.
Language: English
Descriptors: Saccharomyces cerevisiae; Structural genes; Pyruvate decarboxylase; Alleles; Mutants; Promoters; Gene expression; Genetic regulation; Genetic engineering; Enzyme activity; Restriction mapping; Genetic code; Nucleotide sequences; Amino acid sequences
Abstract: Three structural genes encode the pyruvate decarboxylase isoenzymes in the yeast Saccharomyces cerevisiae. PDC1 and PDC5 are active during glucose fermentation where PDC1 is expressed about six times more strongly than PDC5. Expression of PDC6 is weak and seems to be induced in ethanol medium. Consequently, pdc1 delta pdc5 delta double mutants do not ferment glucose and do not grow on glucose medium. Spontaneous mutants, derived from such a pdc1 pdc5 strain, were isolated which could again ferment glucose. They showed pyruvate decarboxylase activity due to a duplication of PDC6. The second copy of PDC6 was expressed under the control of the PDC1 promoter, which was still present in the pdc1 strain. However, the resulting PDC1-PDC6 fusion gene could only partially substitute for PDC1: to achieve normal growth and high pyruvate decarboxylase activity strains carrying PDC1-PDC6 required a functional PDC5 gene which is dispensable in a PDC1 wild-type background. Thus, expression of PDC5 depends on the state of the PDC1 locus: low in the PDC1 wild-type background and high in PDC1-PDC6 fusion strains and, as shown previously, in pdc1 mutants. The activation of PDC5 expression in PDC1-PDC6 strains may be due to particular properties of the PDC1-PDC6 fusion protein or simply to the weaker expression of PDC1-PDC6 in comparison to the wild-type PDC1 gene.
104 NAL Call. No.: QP501.E8
Phenylalanine amonia-lyase in potato (Solanum tuberosum L.).
Genomic complexity, structural comparison of two selected
genes and modes of expression.
Joos, H.J.; Hahlbrock, K.
New York, NY : Springer-Verlag New York Inc; 1992 Mar.
European journal of biochemistry v. 204 (2): p. 621-629; 1992
Mar. Includes references.
Language: English
Descriptors: Solanum tuberosum; Phenylalanine ammonia-lyase; Structural genes; Messenger RNA; Dna; Clones; Genomes; Amino acid sequences; Nucleotide sequences; Phytophthora infestans; Injuries; Stress response; Gene expression
Abstract: Potato (Solanum tuberosum L. cv. Datura) contains approximately 40-50 phenylalanine ammonia-lyase (PAL) genes/haploid genome. Considerable cDNA heterogeneity indicates that at least about 10, and probably more, of these genes are potentially active. One subfamily, represented by one selected member (PAL-1), was analyzed with respect to genomic complexity, nucleotide and deduced amino acid sequence, and mode of constitutive or induced expression. For comparison, a second gene (PAL-2), representing several subfamilies that are easily distinguished from PAL-1, was included in these studies. Extensive structural similarities were observed both between the TATA-proximal portions of the PAL-1 and PAL-2 promoters, particularly in the areas containing putative cis-acting elements, and among all presently known PAL proteins from various higher and lower plants. The relative abundance of PAL mRNA varied greatly in several major potato organs. However, the patterns obtained with probes detecting either total PAL mRNA or more specifically, PAL-1-related or PAL-2-related mRNA species, were the same within experimental error. Mature leaves contained particularly low levels of PAL mRNA. Infection of these leaves with the pathogenic fungus, Phytophthora infestans, resulted in a large, transient induction of PAL mRNA. The relative timing of PAL-1 and PAL-2 mRNA expression, however, differed in compatible (fungus virulent, plant susceptible) but not in incompatible interactions (fungus avirulent, plant resistant). Wounding of leaves caused an extremely rapid and transient induction of both PAL mRNA species.
105 NAL Call. No.: 448.3 J82
Phosphatidylglycerolphosphate synthase expression in
Schizosaccharomyces pombe is regulated by the phospholipid
precursors inositol and choline. Karkhoff-Schweizer, R.R.;
Kelly, B.L.; Greenberg, M.L.
Washington, D.C. : American Society for Microbiology; 1991
Oct. Journal of bacteriology v. 173 (19): p. 6132-6138; 1991
Oct. Includes references.
Language: English
Descriptors: Yeasts; Diphosphatidylglycerols; Ligases; Mitochondria; Membranes; Gene expression; Genetic regulation; Myo-inositol; Choline; Enzyme activity; Kinetics; Temperature
Abstract: The enzyme phosphatidylglycerolphosphate synthase (PGPS; CDP-diacylglycerol glycerol 3-phosphate 3- phosphatidyltransferase; EC 2.7.8.5) catalyzes the committed step in the cardiolipin biosynthetic pathway. To study the regulation of PGPS in Schizosaccharomyces pombe, we characterized the enzyme biochemically. Maximum activity occurred in the presence of 6 mM Triton X-100 at pH 7.5. The apparent Km values for CDP-diacylglycerol and glycerol 3- phosphate were 130 and 26 micromole, respectively. Optimal activity was at 35 degree C, and enzyme activity was labile above 40 degree C. Thioreactive agents were inhibitory to PGPS activity. To determine whether S. pombe PGPS is regulated by phospholipid precursors, we examined the time-dependent expression of PGPS upon inositol and choline starvation. Starvation for inositol resulted in a threefold increase in PGPS expression in wild-type cells. In cho1 and cho2 mutants, which are blocked in phosphatidylcholine synthesis, starvation for choline resulted in transient derepression of PGPS expression. In choline auxotrophs starved for inositol, PGPS was derepressed 2.5- to 3-fold in the presence of choline and less or not at all in the absence of choline. This is the first description of PGPS regulation in S. pombe and the first demonstration of inositol-mediated regulation in the inositolrequiring yeast species.
106 NAL Call. No.: QD341.A2N8
Plant nonsense suppressor tRNATyr gene are expressed at very
low levels in vitro due to inefficient splicing of the introncontaining
pre-tRNAs. Szweykowska-Kulinska, Z.; Beier, H.
Oxford : IRL Press; 1991 Feb25.
Nucleic acids research v. 19 (4): p. 707-712; 1991 Feb25.
Includes references.
Language: English
Descriptors: Arabidopsis thaliana; Nicotiana rustica; Transfer RNA; Genetic regulation; Suppression; Introns
Abstract: Oligonucleotide-directed mutagenesis was used to generate amber, ochre and opal suppressors from cloned Arabidopsis and Nicotiana tRNA(Tyr) genes. The nonsense suppressor tRNA(Tyr) genes were efficiently transcribed in HeLa and yeast nuclear extracts, however, intron excision from all mutant pre-tRNAsTyr was severely impaired in the homologous wheat germ extract as well as in the yeast in vitro splicing system. The change of one nucleotide in the anticodon of suppressor pre-tRNAs leads to a distortion of the potential intron-anticodon interaction. In order to demonstrate that this caused the reduced splicing efficiency, we created a point mutation in the intron of Arabidopsis tRNA(Tyr) which affected the interaction with the wild-type anticodon. As expected, the resulting pre-tRNA was also inefficiently spliced. Another mutation in the intron, which restored the base-pairing between the amber anticodon and the intron of pre-tRNA(Tyr) resulted in an excellent substrate for wheat germ splicing endonuclease. This type of amber suppressor tRNA(Tyr) gene which yields high levels of mature tRNA(Tyr) should be useful for studying suppression in higher plants.
107 NAL Call. No.: 381 B522
Posttranscriptional regulation of the expression of MET2 gene
of Saccharomyces cerevisiae.
Forlani, N.; Martegani, E.; Alberghina, L.
Amsterdam : Elsevier Science Publishers; 1991 May02.
Biochimica et biophysica acta : International journal of
biochemistry and biophysics v. 1089 (1): p. 47-53; 1991 May02.
Includes references.
Language: English
Descriptors: Saccharomyces cerevisiae; Methionine; Biosynthesis; Genetic regulation; Amino acid metabolism; Gene expression
Abstract: The first step of the specific pathway for methionine biosynthesis in the yeast Saccharomyces cerevisiae is catalyzed by the enzyme L-homoserine-O-acetyltransferase (HSTase) (EC 2.3.1.31), encoded by the MET2 gene. In order to ascertain whether there is a posttranscriptional control on the MET2 gene expression, as suggested by previous results on the expression of the cloned gene, systems for high inducible expression of MET2 gene were constructed. In these constructs the MET2 gene was cloned in yeast expression vectors under the control of an inducible yeast GAL promoter element so that the MET2 was transcribed at very high levels under induced conditions. Measurements or the specific mRNA levels showed a strong stimulation of MET2 gene transcription in yeast transformants grown on galactose as carbon source, corresponding to 50-100-fold the repressed conditions, while only a 2-fold increase of the enzymatic activity was observed. In addition, no evidence of a strong induced polypeptide of appropriate size on two dimensional gel electrophoresis was obtained. To understand the functional role of the non-coding 5' region of MET2 mRNA, we performed either a partial and a complete deletion of the 5' leader sequence, but even with these constructs an elevated mRNA level was not associated to a marked increase of the HSTase activity. These data support the idea of a posttranscriptional regulation of MET2 gene expression and show that the untranslated region of the specific mRNA is not involved in this regulatory mechanism.
108 NAL Call. No.: QP601.M49 Probing molecular structure and structural changes of voltagegated channel by expressing mutant channels in yeast and reconstituting them into planar membranes. Colombini, M.; Peng, S.; Blachly-Dyson, E.; Forte, M. San Diego, Calif. : Academic Press; 1992. Methods in enzymology v. 207: p. 432-444; 1992. In the series analytic: Ion channels / edited by B. Rudy and L.E. Iverson. Includes references.
Language: English
Descriptors: Saccharomyces cerevisiae; Membranes; Proteins; Mutants; Mutations; Molecular conformation
109 NAL Call. No.: 381 J8223
Properties and distribution of soybean proglycinin expressed
in Saccharomyces cerevisiae.
Utsumi, S.; Kanamori, J.; Kim, C.S.; Sato, T.; Kito, M.
Washington, D.C. : American Chemical Society; 1991 Jun.
Journal of agricultural and food chemistry v. 39 (6): p.
1179-1186; 1991 Jun. Includes references.
Language: English
Descriptors: Glycine max; Soy protein; Amino acid sequences; Gene expression; Genetic code; Genetic engineering; Immunocytochemistry; Nucleotide sequences; Protein quality; Saccharomyces cerevisiae; Food research
Abstract: Glycinin is a predominant storage protein of soybean. The high-level expression system of preproglycinin cDNA in yeast was established. The signal sequence of the expressed protein was cleaved at the correct site. However, most of the expressed proteins were insoluble due to their interaction with intracellular components at the acidic polypeptide region. The expressed proteins separated from the intracellular components by ion-exchange chromatography in the presence of urea were soluble without urea and self-assembled into trimers. The insolubility of the expressed proteins caused an accumulation of modified proteins with disturbed folding. Immunocytochemical distribution demonstrated that the expressed protein from the cDNA encoding preproglycinin accumulated in Golgi-like structure and organellas which may be derived from Golgi-like structure, and one from the cDNA encoding proglycinin homologue protein was found in the cytosol.
110 NAL Call. No.: QH426.C8 Properties of two nuclear pet mutants affecting expression of the mitochondrial oli1 gene of Saccharomyces cerevisiae. Payne, M.J.; Schweizer, E.; Lukins, H.B. Berlin, W. Ger. : Springer International; 1991. Current genetics v. 19 (5): p. 343-351; 1991. Includes references.
Language: English
Descriptors: Saccharomyces cerevisiae; Mutants; Mitochondria; Gene expression; Atp; Ligases; Temperature; Genetic regulation; Transcription
Abstract: This study details the characteristics of two temperature-conditional pet mutants of yeast, strains ts1860 and ts379, which at the non-permissive temperature show deficiencies in the formation of three mitochondrially encoded subunits of the ATP synthase complex. By analysis of mitochondrial translation products, and of mitochondrial transcription in temperature shift experiments from the permissive (22 degrees C) to the non-permissive (36 degrees C) temperature, it was concluded that the nuclear mutations in both mutants primarily inhibit synthesis of ATP synthase subunit 9, and that reductions in subunit 8 and 6 synthesis are secondary pleiotropic effects. Following transfer to 36 degree C, cells of mutant ts379 display a near complete inhibition of subunit 9 synthesis within 1 h, coincident with a marked reduction in the level of the cognate oli1 mRNA. On the other hand, near complete inhibition of subunit 9 synthesis in strain ts1860 occurs after 3 h at 36 degrees C, at which time there is little change in the level of subunit 9 mRNA. In both mutants the mRNA levels for subunits 6 and 8 are not significantly affected at the time of inhibition of subunit 9 synthesis. Provision of an alternative source of subunit 8, translated extra-mitochondrially for import into the organelle, does not overcome the mutant phenotype of either mutant at 36 degrees C, confirming that subunit 8 is not the sole or primary deficiency in each mutant. The mutants indicate that the products of a least two nuclear genes (designated AEP1 and AEP2) are required for the expression of the mitochondrial oli1 gene and the synthesis of subunit 9. The product of the AEP1 gene (defective in mutant tsl860) is required for translation of oli1 mRNA while the AEP2 product (defective in mutant ts379) is essential either for the stability of oli1 mRNA or for the correct processing of precursor transcripts to the mature message.
111 NAL Call. No.: QR53.B56
Proteolytic degradation of heterologous proteins expressed in
Aspergillus niger.
Archer, D.B.; MacKenzie, D.A.; Jeenes, D.J.; Roberts, I.N.
Middlesex : Science and Technology Letters; 1992 May.
Biotechnology letters v. 14 (5): p. 357-362; 1992 May.
Includes references.
Language: English
Descriptors: Aspergillus niger; Proteinases; Enzyme activity; Proteolysis; Lysozyme; Phospholipase a2; Protein secretion; Genetic transformation
Abstract: Proteolytic degradation of heterologous proteins expressed in the filamentous fungus Aspergillus niger reduces the yield of authentic target protein. The activities of A. niger proteases are differentiated by their effects on two proteins expressed and secreted from A. niger: hen egg-white lysozyme and porcine pancreatic phospholipase A2.
112 NAL Call. No.: QP501.E8
Purification, characterisation and mutagenesis of highly
expressed recombinant yeast pyruvate kinase.
Murcott, T.H.L.; McNally, T.; Allen, S.C.; Fothergill-Gilmore,
L.A.; Muirhead, H.
Secaucus, N.J. : Springer-Verlag New York Inc; 1991 Jun.
European journal of biochemistry v. 198 (2): p. 513-519; 1991
Jun. Includes references.
Language: English
Descriptors: Saccharomyces cerevisiae; Pyruvate kinase; Recombination; Gene expression; Purification; Characterization; Mutagenesis
Abstract: Recombinant yeast pyruvate kinase has been purified from a strain of Saccharomyces cerevisiae expressing the enzyme to very high levels. Expression was from a multicopy plasmid under the control of the yeast phosphoglycerate kinase promoter. The gene was expressed in the absence of the genomically encoded pyruvate kinase, using a strain of yeast in which the pyruvate kinase gene has been disrupted by the insertion of the yeast Ura3 gene. The purification procedure minimised proteolytic artefacts and enabled the covenient purification of 15-20 mg enzyme from 1 l culture. The purified enzyme was characterised by a high specific activity and by a lack of proteolytic degradation. Two active-site mutants of yeast pyruvate kinase have been produced, expressed and characterised in this system and preliminary results are described.
113 NAL Call. No.: 500 N21P
Regulated expression of the GAL4 activator gene in yeast
provides a sensitive genetic switch for glucose repression.
Griggs, D.W.; Johnston, M.
Washington, D.C. : The Academy; 1991 Oct01.
Proceedings of the National Academy of Sciences of the United
States of America v. 88 (19): p. 8597-8601; 1991 Oct01.
Includes references.
Language: English
Descriptors: Saccharomyces cerevisiae; Strains; Yeasts; Gene expression; Genetic regulation; Glucose; Inhibitors; Mutations; Promoters; Synergism; Transcription
Abstract: Glucose (catabolite) repression is mediated by multiple mechanisms that combine to regulate transcription of the GAL genes over at least a thousandfold range. We have determined that this is due predominantly to modest glucose repression (4- to 7-fold) of expression of GAL4, the gene encoding the transcriptional activator of the GAL genes. GAL4 regulation is affected by mutations in several genes previously implicated in the glucose repression pathway; it is not dependent on GALA or GAL80 protein function. GAL4 promoter sequences that mediate glucose repression were found to lie downstream of positively acting elements that may be "TATA boxes." Two nearly identical sequences (10/12 base pairs) in this region that may be binding sites for the MIG1 protein were identified as functional glucose-control elements. A 4- base-pair insertion in one of these sites causes constitutive GAL4 synthesis and leads to substantial relief (50-fold) of glucose repression of GAL1 expression. Furthermore, promoter deletions that modestly reduce GAL4 expression, and therefore presumably the amount of GAL4 protein synthesized, cause much greater reductions in GAL1 expression. These results suggest that GAL4 works synergistically to activate GAL1 expression. Thus, glucose repression of GAL1 expression is due largely to a relatively small reduction of GAL4 protein levels caused by reduced GAL4 transcription. This illustrates how modest regulation of a weakly expressed regulatory gene can act as a sensitive genetic switch to produce greatly amplified responses to environmental changes.
114 NAL Call. No.: 381 B523
Rotation and interactions of genetically expressed cytochrome
P-450IA1 and NADPH-cytochrome P-450 reductase in yeast
microsomes.
Iwase, T.; Sakaki, T.; Yabusaki, Y.; Ohkawa, H.; Ohta, Y.;
Kawato, S. Washington, D.C. : American Chemical Society; 1991
Aug27. Biochemistry v. 30 (34): p. 8347-8351; 1991 Aug27.
Includes references.
Language: English
Descriptors: Saccharomyces cerevisiae; Microsomes; Cytochrome p-450; Nadph-cytochrome-c2 reductase; Gene expression; Vectors; Spectral analysis; Rats
Abstract: Rat liver cytochrome P-450IA1 and/or yeast NADPHcytochrome P-450 reductase was expressed genetically in yeast microsomes. The ratio of P-45OIA1 to the reductase was about 17:1 and 1:2 without and with coexpression of the reductase, respectively. Rotational diffusion of P-450IA1 was examined by observing the flash-induced absorption anisotropy, r(t), of the heme-CO complex. In only P-450Ia1-expressed microsomes, 28% of P-450IA1 was rotating with a rotational relaxation time of about 1200 microseconds. The mobile population was increased to 43% by the presence of the coexpressed reductase, while rotational relaxation time was not changed significantly. Increased concentration of KCl from 0 to 1000 mM caused considerable mobilization of P-450IA1. The results demonstrate a proper incorporation of P-450IA1 molecules into yeast microsomal membranes. The significant mobilization of P-450IA1 by the presence of reductase suggests a possible transient association of P-450IA1 with the reductase.
115 NAL Call. No.: 381 J824
Saccharomyces cerevisiae elongation factor 2: genetic cloning,
characterization of expression, and G-domain modeling.
Perentesis, J.P.; Phan, L.D.; Gleason, W.B.; LaPorte, D.C.;
Livingston, D.M.; Bodley, J.W.
Baltimore, Md. : American Society for Biochemistry and
Molecular Biology; 1992 Jan15.
The Journal of biological chemistry v. 267 (2): p. 1190-1197;
1992 Jan15. Includes references.
Language: English
Descriptors: Saccharomyces cerevisiae; Structural genes; Cloning; Gene expression; Nucleotide sequences; Amino acid sequences
Abstract: The elongation factor 2 (EF-2) genes of the yeast Saccharomyces cerevisiae have been cloned and characterized with the ultimate goal of gaining a better understanding of the mechanism and control of protein synthesis. Two genes (EFT1 and EFT2) were isolated by screening a bacteriophage lambda yeast genomic DNA library with an oligonucleotide probe complementary to the domain of EF-2 that contains diphthamide, the unique posttranslationally modified histidine that is specifically ADP-ribosylated by diphtheria toxin. Although EFT1 and EFT2 are located on separate chromosomes, the DNA sequences of the two genes differ at only four positions out of 2526 base pairs, and the predicted protein sequences are identical. Genetic deletion of each gene revealed that at least one functional copy of either EFT gene is required for cell viability. Messenger RNA levels of yeast EF-2 parallel cellular growth and peak in mid-log phase cultures. The EF-2 protein sequence is strikingly conserved through evolution. Yeast EF-2 is 66% identical to, and shares over 85% homology with, human EF-2. In addition, yeast and mammalian EF-2 share identical sequences at two critical functional sites: (i) the domain containing the histidine residue that is modified to diphthamide and (ii) the threonine residue that is specifically phosphorylated in vivo in mammalian cells by calmodulin-dependent protein kinase III, also known as EF-2 kinase. Furthermore, yeast EF-2 also contains the Glu-X-X-ArgX -Ile-Thr-Ile "effector" sequence motif that is conserved among all known elongation factors, and its GTP-binding domain exhibits strong homology to the G-domain of Escherichia coli elongation factor Tu (EF-TU) and other G-protein family members. Based upon these observations, we have modeled the Gdomain of the deduced EF-2 protein sequence to the solved crystallographic structure for EF-TU.
116 NAL Call. No.: 442.8 Z34
The Saccharomyces cerevisiae SPT14 gene is essential for
normal expression of the yeast transposon, Ty, as well as for
expression of the HIS4 gene and several genes in the mating
pathway.
Fassler, J.S.; Gray, W.; Lee, J.P.; Yu, G.; Gingerich, G.
Berlin, W. Ger. : Springer International; 1991 Nov.
M G G : Molecular and general genetics v. 230 (1/2): p.
310-320; 1991 Nov. Includes references.
Language: English
Descriptors: Saccharomyces cerevisiae; Transposable elements; Genes; Dna binding proteins; Nucleotide sequences; Gene expression; Transcription; Genetic regulation; Mating; Gene mapping; Gene location; Chromosomes; Restriction mapping; Messenger RNA; Mutations
Abstract: To investigate the role of the trans-acting transcription factor encoded by the essential SPT14 (SPT=suppressor of Ty insertion mutations) gene, we have cloned, mapped and sequenced the gene. From the analysis of the effect of spt14 mutations on expression of various genes, we conclude that the SPT14 product has an important role in activation of Ty transcription as well as in the regulation of other genes including HIS4 and several of the a- and alphaspecific mating type genes. Similarities in the phenotypes of spt14 and spt13 mutants (suppression of Ty insertion mutations but not delta insertion mutations), lead to the suggestion that the SPT14 gene and the previously characterized SPT13/GAL11 gene might encode transcriptional regulators with related functions. Our current findings show that in contrast to SPT13/GAL11, which appears negatively to regulate Ty transcription, SPT14 plays a role in the activation of Ty transcription. Thus, despite the similarities in the suppression phenotype exhibited by spt13 and spt14 mutants, SPT13/GAL11 and SPT14 probably differ in their transcriptional roles.
117 NAL Call. No.: 500 N21P
Selective induction of gene expression and second-messenger
accumulation in Dictyostelium discoideum by the partial
chemotactic antagonist 8-p-chlorophenylthioadenosine 3',5'-
cyclic monophosphate. Peters, D.J.M.; Bominaar, A.A.; SnaarJagalska,
B.E.; Brandt, R.; Haastert, P.J.M. van; Ceccarelli,
A.; Williams, J.G.; Schaap, P.
Washington, D.C. : The Academy; 1991 Oct15.
Proceedings of the National Academy of Sciences of the United
States of America v. 88 (20): p. 9219-9223. ill; 1991 Oct15.
Includes references.
Language: English
Descriptors: Dictyostelium; Molds; Strains; Binding proteins; C-amp; Gene expression; Genetic regulation; Phospholipids; Inositol phosphates
Abstract: During development of the cellular slime mold Dictyostelium discoideum, cAMP induces chemotaxis and expression of different classes of genes by means of interaction with surface cAMP receptors. We describe a cAMP derivative, 8-p-chlorophenylthioadenosine 3',5'-cyclic monophosphate (8-CPT-cAMP), which inhibits cAMP-induced chemotaxis at low concentrations but induces chemotaxis at supersaturating concentrations. This compound, moreover, selectively activates expression of aggregative genes but not of postaggregative genes. 8-CPT-cAMP induces normal cGMP and cAMP accumulation but in contrast to cAMP, which increases inositol 1,4,5-trisphosphate levels, 8-CPT-cAMP decreases inositol 1,4,5-trisphosphate levels. The derivative induces reduced activation of guanine nucleotide regulatory proteins, which may cause its defective activation of inositol 1,4,5- trisphosphate production. Our data suggest that disruption of inositolphospholipid signaling impairs chemotaxis and expression of a subclass of cAMP regulated genes.
118 NAL Call. No.: QH426.C8
Sequence analysis of the ARG7 gene of Schizosaccharomyces
pombe coding for argininosuccinate lyase. Expression of the
gene in Saccharomyces cerevisiae. Loppes, R.; Michels, R.;
Decroupette, I.; Joris, B.
Berlin, W. Ger. : Springer International; 1991.
Current genetics v. 19 (4): p. 255-260; 1991. Includes
references.
Language: English
Descriptors: Yeasts; Saccharomyces cerevisiae; Genetic code; Nucleotide sequences; Amino acid sequences; Argininosuccinate lyase; Enzyme activity; Gene expression; Gene mapping
119 NAL Call. No.: 442.8 G28
Site-specific recombination determined by I-SceI, a
mitochondrial group I intron-encoded endonuclease expressed in
the yeast nucleus. Plessis, A.; Perrin, A.; Haber, J.E.;
Dujon, B.
Baltimore, Md. : Genetics Society of America; 1992 Mar.
Genetics v. 130 (3): p. 451-460; 1992 Mar. Includes
references.
Language: English
Descriptors: Saccharomyces cerevisiae; Introns; Mitochondrial DNA; Nucleases; Gene expression; Nuclei; Recombination; Dna modification; Enzyme activity; Dna repair; Gene conversion; Deletions
Abstract: The Saccharomyces cerevisiae mitochondrial endonuclease I-SceI creates a double-strand break as the initiating step in the gene conversional transfer of the omega+ intron to omega- DNA. We have expressed a galactoseinducible synthetic I-SceI gene in the nucleus of yeast that also carries the I-SceI recognition site on a plasmid substrate. We find that the galactose-induced I-SceI protein can be active in the nucleus and efficiently catalyze recombination. With a target plasmid containing direct repeats of the Escherichia coli lacZ gene, one copy of which is interrupted by a 24-bp cutting site, galactose induction produces both deletions and gene conversions. Both the kinetics and the proportion of deletions and gene conversions are very similar to analogous events initiated by a galactoseinducible HO endonuclease gene. We also find that, in a rad52 mutant strain, the repair of double-strand breaks initiated by I-SceI and by HO are similarly affected: the formation of deletions is reduced, but not eliminated. Altogether, these results suggest either that the two endonucleases act in the same way after double-strand break formation or that the two endonucleases are not involved in subsequent steps.
120 NAL Call. No.: 450 P692
Slow-growth phenotype of transgenic tomato expressing
apoplastic invertase. Dickinson, C.D.; Altabella, T.;
Chrispeels, M.J.
Rockville, Md. : American Society of Plant Physiologists; 1991
Feb. Plant physiology v. 95 (2): p. 420-425. ill; 1991 Feb.
Includes references.
Language: English
Descriptors: Lycopersicon esculentum; Genetic transformation; Transgenics; Gene expression; Beta-fructofuranosidase; Phenotypes; Growth; Inhibition; Cauliflower mosaic caulimovirus; Symptoms; Light; Photosynthates; Translocation
Abstract: The growth of transgenic tomato (Lycopersicon esculentum) plants that express in their apoplast yeast invertase under the control of the cauliflower mosaic virus 35S promoter is severely inhibited. The higher the level of invertase, the greater the inhibition of growth. A second phenotypic characteristic of these transgenic plants is the development of yellow and necrotic spots on the leaves, and leaf curling. Again the severity of the symptoms is correlated with the level of invertase. These symptoms do not develop in shaded leaves indicating the need for photosynthesis. Keeping the plants in the dark for a prolonged period (24 hours) results in the disappearance of leaf starch from the control plants, but not from the plants with apoplastic invertase. These results are consistent with the interpretation that apoplastic invertase prevents photosynthate export from source leaves and that phloem loading includes an apoplastic step.
121 NAL Call. No.: QH426.C8
Structure and express of a plastid-encoded groEL homologous
heat-shock gene in a thermophilic unicellular red alga.
Maid, U.; Steinmuller, R.; Zetsche, K.
Berlin, W. Ger. : Springer International; 1992.
Current genetics v. 21 (6): p. 521-525; 1992. Includes
references.
Language: English
Descriptors: Rhodophyta; Chloroplast genetics; Plastids; Dna; Genes; Plant proteins; Nucleotide sequences; Amino acid sequences; Transcription; Gene expression; Translation; Heat shock; Restriction mapping
Abstract: A gene homologous to the E. coli groEL locus was identified on the plastid genome of the unicellular red alga Cyanidium caldarium strain 14-1-1 (synonym: Galdieria sulphuraria). The complete nucleotide sequence was determined and compared to bacterial- and nuclear-encoded counterparts of higher plants. At the amino-acid level the C. caldarium gene shows 70% homology to the corresponding gene of the cyanobacterium Synechococcus and 52% homology to nuclearencoded counterparts of higher plants, respectively. Northern and Western blot experiments were used to investigate the dependence of the transcript- and protein-level on culture temperature and heat shock.
122 NAL Call. No.: 472 N21 SWI6 protein is required for transcription of the periodically expressed DNA synthesis genes in budding yeast. Lowndes, N.F.; Johnson, A.L.; Breeden, L.; Johnston, L.H. London : Macmillan Magazines Ltd; 1992 Jun11. Nature v. 357 (6378): p. 505-508; 1992 Jun11. Includes references.
Language: English
Descriptors: Yeasts; Dna; Gene expression; Proteins; Transcription
Abstract: IN budding yeast many genes are expressed under cell-cycle control in late G1. These include a large group of DNA synthesis genes, the HO gene involved in mating-type switching, C7S1 (chitinase) and also CLN1 and CLN2 (ref. 4) encoding G1 cyclins. Two factors, encoded by the SW14 and SW16 genes, are required for HO (ref. 5), CLN (refs 6, 7) and CTS1 (ref. 3) gene expression and, at least in the HO promoter, bind to CACGA4 upstream sequences (CCBs). This motif is not found upstream of the DNA synthesis genes, which instead have a hexamer element, ACGCGT (MCB), an MluI restriction site, that is recognized by a cell-cycle regulated transcription complex DSC1 (ref. 1). This MluI-activation system consisting of the MCBs and DSC1 is conserved in fission yeast where a DSC1-like complex controls the cdc22+ ribonucleotide reductase gene. The Schizosacharomyces pombe cdc10+ gene encodes a component of DSC1 (ref. 10) and, significantly, this has homology with both the Swi4 and Swi6 proteins. Here we show that Swi6 is an essential component of DSC1 and that deletion of SW16 impairs the cell-cycle regulation of the DNA synthesis genes, as well as CLN1 and CLN2. Thus Swi6 is the common factor in regulation of all the above genes and may therefore be responsible for the timing of their expression in late G1.
123 NAL Call. No.: QH442.A1G4 Synthesis and expression of gene encoding tuna, pigeon, and horse cytochromes c in the yeast Saccharomyces cerevisiae. Hickey, D.R.; Jayaraman, K.; Goodhue, C.T.; Shah, J.; Fingar, S.A.; Clements, J.M.; Hosokawa, Y.; Tsunasawa, S.; Sherman, F. Amsterdam : Elsevier Science Publishers; 1991. Gene v. 105 (1): p. 73-81; 1991. Includes references.
Language: English
Descriptors: Saccharomyces cerevisiae; Horses; Thunnus; Pigeons; Synthetic genes; Cytochrome c; Gene expression; Messenger RNA; Vectors; Cloning; Enzyme activity; Nucleotide sequences; Amino acid sequences
Abstract: Genes encoding tuna, pigeon, and horse cytochromes c- were constructed with synthetic oligodeoxyribonucleotides having preferred codons and portions of the iso-1-cytochrome c-encoding gene from the yeast Saccharomyces cerevisiae. The genes were ligated into an expression vector, which contains the normal 5'- and 3'-untranslated regions of the yeast iso-1- cytochrome c gene, and were integrated in single copy into the chromosome. Yeast strains were also constructed with multiple integrated copies of the pigeon gene. The heterologous and normal mRNA levels of the single-copy strains were equivalent. Although the N-terminal methionines were completely cleaved in the heterospecific proteins, the levels of trimethylation of Lys72 and acetylation of N-terminal glycines ranged from 39-78% and 10-70%, respectively. Horse cytochrome c was produced at a nearly normal level, whereas the pigeon and tuna cytochromes c were produced at approx. 40% of the normal levels. The levels of the cytochromes c and growth of the mutant yeast strains indicated that the heterospecific cytochromes had approx. 50% specific activity in vivo.
124 NAL Call. No.: 500 N484
Systems and approaches for expression and secretion of
heterologous proteins in the filamentous fungus Aspergillus
niger var. awamori: current status. Berka, R.M.
New York, N.Y. : The Academy; 1991.
Annals of the New York Academy of Sciences v. 646: p. 207-211;
1991. In the series analytic: Recombinant DNA technology I /
edited by A. Prokop and R.K. Bajpai. Includes references.
Language: English
Descriptors: Aspergillus niger; Chymosin; Fungal protein; Gene expression; Genetic engineering; Food industry; Industrial microbiology
125 NAL Call. No.: SB599.P45
Temperature and genotype interactions in the expression of
host resistance in lettuce downy mildew.
Judelson, H.S.; Michelmore, R.W.
London : Academic Press; 1992 Apr.
Physiological and molecular plant pathology v. 40 (4): p.
233-245; 1992 Apr. Includes references.
Language: English
Descriptors: Lactuca sativa; Bremia lactucae; Host specificity; Disease resistance; Genotypes; Gene expression; Virulence; Genes; Environmental factors; Air temperature; Genotype environment interaction
Abstract: The influence of temperature and genotype on infection types determined by 13 gene-for-gene interactions between lettuce and the downy mildew fungus, Bremia lactucae, was evaluated. Lettuce seedlings containing each of 13 resistance (Dm) genes were inoculated with isolates of B. lactucae expressing various combinations of the matching avirulence genes, and incubated at 5, 10, 15, or 22 degrees C. Resistance, assessed by the absence of sporulation, became less effective at lower temperatures for Dm6, Dm7, Dm11, Dm15, and Dm16. No change was observed for resistance conditioned by eight other gene-for-gene interactions. Histological examination indicated that the extent of fungal development in host tissue was dependent on the interacting genotypes and on temperature. Manipulation of interactions involving Dm7 and Dm11, using temperature-shift incubation regimes, indicated that continued and cumulative exposure to permissive temperatures was required for effective resistance and suggested that the determinants of specificity are present in most host cells and are expressed throughout fungal development.
126 NAL Call. No.: 448.3 J823
The thn mutation of Schizophyllum commune, which suppresses
formation aerial hyphae, affects expression of the Sc3
hydrophobin gene.
Wessels, J.G.H.; Vries, O.M.H. de; Asgeirsdottir, S.A.;
Springer, J. Reading : Society for General Microbiology; 1991
Oct.
The Journal of general microbiology v. 137 (pt.10): p.
2439-2445; 1991 Oct. Includes references.
Language: English
Descriptors: Schizophyllum commune; Genes; Mutants; Mutations; Proteins; Hyphae; Fungal morphology; Fruiting; Phenotypes; Suppression; Gene expression
Abstract: The spontaneous and recessive mutation thn in the basidiomycete Schizophyllum commune suppresses the formation of aerial hyphae in the monokaryon and, if present as a double dose, the formation of both aerial hyphae and fruit-bodies in the dikaryon. In the monokaryon, the mutation prevents accumulation of mRNA of the Sc3 gene, and in the dikaryon it also prevents the accumulation of fruiting-specific mRNAs, including mRNAs of the Sc1 and Sc4 genes, which are homologous to the Sc3 gene. These three genes code for hydrophobins, a family of small hydrophobic cysteine-rich proteins. In the thn monokaryon, the only detectable change in synthesized proteins is the disappearance of an abundant protein of apparent Mr = 28 K from the culture medium and from the cell walls. Protein sequencing shows that this is the product of the Sc3 gene. The Sc3 hydrophobin is present in the walls of aerial hyphae as a hot-SDS-insoluble complex. Submerged hyphae excrete large amounts of the hydrophobin into the medium.
127 NAL Call. No.: QD341.A2N8
Transcript mapping reveals different expression strategies for
the bicistronic RNAs of the geminivirus wheat dwarf virus.
Dekker, E.L.; Woolston, C.J.; Xue, Y.; Cox, B.; Mullineaux,
P.M. Oxford : IRL Press; 1991 Aug11.
Nucleic acids research v. 19 (5): p. 4075-4081; 1991 Aug11.
Includes references.
Language: English
Descriptors: Wheat dwarf geminivirus; Transcription; Genetic regulation; Gene expression; Rna; Polymerase chain reaction; Molecular mapping; Nucleotide sequences
Abstract: We have characterised the major transcripts of the Czech isolate of wheat dwarf virus (WDV-CJI) which show that WDV uses two different mechanisms for expressing overlapping open reading frames (ORFs). Mapping of the virion sense RNAs identified a single polyadenylated transcript of 1.1kb spanning the overlapping ORFs V1 and V2 which encode cell-cell spread functions and the coat protein respectively. This finding distinguishes WDV from other monocot-infecting geminiviruses studied so far which were shown to encode two 3' co-terminal transcripts capable of expressing either the V1 or V2 ORF. A survey of codon usage at the junction between the V1 and V2 ORF has led us to propose that translational frame shifting analogous to that in the yeast Ty element may occur. Analysis of polymerase chain reaction (PCR) amplified complementary sense cDNA clones has revealed the presence of mature spliced and unspliced RNAs which could encode products of an intron mediated C1:C2 ORF fusion or the C1 ORF product alone. Mapping of the 51 and 31 extremities of the major WDV encoded transcripts has allowed us to identify putative transcription regulatory sequences and the presence of multiple overlapping transcripts may suggest temporal regulation of transcription.
128 NAL Call. No.: QH426.C8
Transient expression of firefly luciferase in protoplasts of
the green alga Chlorella ellipsoidea.
Jarvis, E.E.; Brown, L.M.
Berlin, W. Ger. : Springer International; 1991.
Current genetics v. 19 (4): p. 317-321; 1991. Includes
references.
Language: English
Descriptors: Chlorella ellipsoidea; Protoplasts; Genetic transformation; Luciferase; Reporter genes; Gene expression; Gene mapping
129 NAL Call. No.: QH426.C8
Transient expression of genes in the oomycete Phytophthora
infestans using Bremia lactucae regulatory sequences.
Judelson, H.S.; Michelmore, R.W.
Berlin, W. Ger. : Springer International; 1991.
Current genetics v. 19 (6): p. 453-459; 1991. Includes
references.
Language: English
Descriptors: Bremia lactucae; Phytophthora infestans; Genetic transformation; Gene expression; Beta-glucuronidase; Reporter genes; Vectors; Plant pathogenic fungi
Abstract: Vectors containing fusions between the reporter gene beta-glucuronidase (GUS) and transcriptional regulatory signals from the ham34 and hsp70 genes of the oomycete pathogen, Bremia lactucae, were introduced into protoplasts of the related fungus Phytophthora infestans. Transient expression of the GUS gene was detected when DNA was introduced into protoplasts of P. infestans by treatments with polyethylene glycol and CaCl(2), with cationic liposomes, or by electroporation. After optimization of each procedure, using the transient expression assays, cationic liposomes were identified as the superior method for DNA uptake. Vectors containing the 5'hsp70 sequences and 3'ham34 sequences resulted in the maximum level of GUS activity. The identification of functional vectors for transformation, and of optimal methods for introducing DNA, should assist in achieving stable transformation of P. infestans and other oomycete the fungi.
130 NAL Call. No.: 464.8 P56
Transient expression of the beta-glucuronidase gene introduced
into Uromyces appendiculatus uredospores by particle
bombardment.
Bhairi, S.M.; Staples, R.C.
St. Paul, Minn. : American Phytopathological Society; 1992
Sep. Phytopathology v. 82 (9): p. 986-989; 1992 Sep. Includes
references.
Language: English
Descriptors: Phaseolus vulgaris; Uromyces appendiculatus; Infectivity; Genetic regulation; Gene expression; Genetic transformation; Beta-glucuronidase
Abstract: Plasmids carrying the beta-glucuronidase gene (GUS) under the control of the promoter from the previously cloned gene specific for infection structure, INF24, were constructed by inserting the INF24 promoter in front of the GUS gene carried in the Bluescript vector. These plasmid DNAs were then introduced into uredospores of the bean rust fungus, Uromyces appendiculatus; the biolistic particle delivery system was used. With germination and differentiation on collodion membranes, GUS activity assayed histochemically with X-gluc was evident only in those germlings that were bombarded with the plasmid containing the entire promoter region; it was not evident in those that were mock-bombarded or bombarded with the plasmid lacking the promoter.
131 NAL Call. No.: QH442.A1G4 A
twin-reporter vector for simultaneous analysis of expression
signals of divergently transcribed, contiguous genes in
filamentous fungi. Punt, P.J.; Greaves, P.A.; Kuyvenhoven, A.;
Deutekom, J.C.T. van; Kinghorn, J.R.; Pouwels, P.H.; Hondel,
C.A.M.J.J. van
Amsterdam : Elsevier Science Publishers; 1991.
Gene v. 104 (1): p. 119-122; 1991. Includes references.
Language: English
Descriptors: Aspergillus nidulans; Recombinant DNA; Genes; Vectors; Beta-galactosidase; Beta-glucuronidase; Gene expression; Nitrate reductase; Nitrite reductase
Abstract: To analyze the promoter region(s) of divergently. Transcribed fungal genes, a twin-reporter vector was constructed. This vector contains two divergently oriented reported genes, encoding Escherichia coli beta-glucuronidase (uidA) and E. coli beta-galactosidase (lacZ). Terminator regions of the Aspergillus nidulans nitrate and nitrite reductase-encoding genes, niaD and niiA, respectively, have been cloned 3' to the reporter genes to ensure proficient transcription termination of the reporter genes. The reporter genes have been separated by a unique NotI restriction site, which can be used for the insertion of expression signals. A mutant argB selection marker has been introduced in order to obtain A. nidulans transformants with a single copy of the vector integrated at the argB locus. The use of the vector was demonstrated by insertion of the A. nidulans niaD-niiA intergenic region and analysis of A. nidulans transformants obtained with this construct. Control of expression of both reporter genes was found to be in accordance with previously published data on control of nitrate assimilation [Cove, Biol. Rev. 54 (1979) 291-327].
132 NAL Call. No.: 381 J824
Two yeast genes encoding calmodulin-dependent protein kinases.
Isolation, sequencing, and bacterial expressions of CMK1 and
CMK2.
Ohya, Y.; Kawasaki, H.; Suzuki, K.; Londesborough, J.; Anraku,
Y. Baltimore, Md. : American Society for Biochemistry and
Molecular Biology; 1991 Jul05.
The Journal of biological chemistry v. 266 (19): p.
12784-12794; 1991 Jul05. Includes references.
Language: English
Descriptors: Saccharomyces cerevisiae; Protein kinase; Calmodulin; Gene expression; Cloning; Structural genes; Isolation; Nucleotide sequences; Amino acid sequences
Abstract: We have isolated two genes from Saccharomyces cerevisiae that both encode a calmodulin-dependent protein kinase (CaM kinase). The CMK1 gene has been cloned by hybridization using an oligonucleotide probe synthesized on the basis of the peptide sequence of purified yeast CaM kinase (Londesborough, J. (1989) J. Gen. Microbiol. 135, 3373-3383). The other gene, CMK2, which is homologous to CMK1, has been isolated by screening at low stringency with a CMK1 fragment as a probe. The CMK2 product expressed in bacteria shows Ca2+- and CaM-dependent protein kinase activity, indicating that CMK2 also encodes a CaM kinase. The CMK1 and CMK2 products expressed in bacteria were found to have different biochemical properties in terms of autoregulatory activity and preference for yeast CaM or bovine CaM for maximal activity. Antibody raised against a peptide fragment of the CMK1 protein crossreacts with the CMK2 product. Immunoblotting with this antibody indicated that the CMK1 and CMK2 products have apparent molecular masses of 56 and 50 kDa, respectively, in yeast cells. The predicted amino acid sequences of the two CMK products exhibit highest similarity with mammalian calmodulindependent multifunctional protein kinase II (CaM kinase II): the similarity within the N-terminal catalytic domain is about 40%, whereas that within the rest of the sequence is 25%. These data indicate that yeast has two kinds of genes encoding CaM kinase isozymes whose structural and functional properties are closely related to those of mammalian caM kinase II. Another gene may be substituted for function of the CMK1 and CMK2 kinase in vivo, since elimination of both kinase genes is not lethal.
133 NAL Call. No.: QH426.C8 Ubiquitin gene expression: response to environmental changes. Fraser, J.; Luu, H.A.; Neculcea, J.; Thomas, D.Y.; Storms, R.K. Berlin, W. Ger. : Springer International; 1991. Current genetics v. 20 (1/2): p. 17-23; 1991. Includes references.
Language: English
Descriptors: Saccharomyces cerevisiae; Gene expression; Plant proteins; Genes; Transcription; Messenger RNA; Genetic regulation; Heat shock; Methyl methanesulfonate; Mutagens; Starvation; Refeeding
Abstract: It has previously been shown that the yeast ubiquitin genes UBI1, 2 and 3 are strongly expressed during the log-phase of batch culture growth, whereas the U8I4 gene is weakly expressed. We found that heat shock, treatment with DNA-damaging agents, starvation, and the feeding of starved cells all transiently induced UBI4. These results suggest that UBI4 is induced whenever a change in culture conditions dictates a dramatic shift in cellular metabolism, and that UBI4 expression returns to lower levels once cellular metabolism has adapted to the new conditions. In contrast, all of the treatments tested, except starvation, transiently repressed the UBI1, 2 and 3 genes. Although starvation also repressed UBI1, 2 and 3 its effect was not transient, and expression only recovered upon the addition of fresh media. These results, together with others presented here, suggest that high levels of UBI1, 2 and 3 expression are dependant upon ongoing cell growth, and that treatments which slow or stop growth repress their expression.
134 NAL Call. No.: QH442.A1G4
Vectors for the expression and analysis of DNA-binding
proteins in yeast. Bonner, J.J.
Amsterdam : Elsevier Science Publishers; 1991.
Gene v. 104 (1): p. 113-118; 1991. Includes references.
Language: English
Descriptors: Saccharomyces cerevisiae; Recombinant DNA; Plasmids; Heat shock; Binding proteins; Nucleotide sequences; Vectors
Abstract: A series of 13 vectors is described. All are yeast centromere plasmids with the LEU2 gene for selection in yeast, and pUC19 sequences for growth in Escherichia coli. All contain the GAL1 promoter directing transcription into a multiple cloning site (MCS). For twelve of the plasmids, synthetic oligodeoxyribonucleotides create an ATG start codon, in a productive context for yeast, prior to the MCS. Spacing between the ATG and the MCS is variable, to facilitate the cloning of gene fragments in the appropriate reading frame. Nine of the plasmids also contain the strong transcriptional activator from the herpes simplex virus VP16 gene, joined downstream from the MCS. In these nine vectors, all possible combinations of reading frames are available. The suitability of these plasmids for the expression and analysis of DNAbinding domains is tested by cloning into them fragments of the yeast HSF1 gene, encoding the heat shock transcription factor (HSF). The regulation of reporter gene expression by the chimeric HSF-VP16 fusions is described, as is the utility of these vectors for other applications.
135 NAL Call. No.: QH506.E46
Virus-like genetic organization and expression strategy for a
double-stranded RNA genetic element associated with biological
control of chestnut blight. Shapira, R.; Choi, G.H.; Nuss,
D.L.
Oxford, Eng. : IRL Press; 1991 Apr.
The EMBO journal - European Molecular Biology Organization v.
10 (4): p. 731-739; 1991 Apr. Includes references.
Language: English
Descriptors: Castanea sativa; Cryphonectria parasitica; Blight; Plant viruses; Plant disease control; Biological control; Hypovirulence; Rna; Nucleotide sequences
Abstract: The complete nucleotide sequence of the largest double-stranded (ds) RNA present in hypovirulent strain EP713 of the chestnut blight pathogen, Cryphonectria parasitica, was determined and the predicted genetic organization was confirmed by translational mapping analysis. The deduced RNA sequence was 12 712 bp in length, excluding the terminal poly(A):poly(U) homopolymer domain. The strand terminating with 3'-poly(A) contained two contiguous large open reading frames (ORF A and ORF B) beginning at nucleotide residues 496-498 and extending to nucleotide positions 11 859-11 861. The junction between ORF A and ORF B consisted of the sequence 5'-UAAUG-3', where UAA served as the termination codon for ORF A and AUG was the 5'-proximal initiation codon within ORE B. ORF A (622 codons in length, excluding the termination codon) was recently shown to encode two polypeptides, p29 and p40, which were generated from a nascent polyprotein by an autocatalytic event mediated by p29 (Choi et al., 1991). A similar autocatalytic event was observed during in vitro translation of ORF B (3165 codons in length) resulting in the release of a 48 kd polypeptide from the amino-terminal portion of the ORF B-encoded polyprotein. These results are discussed in terms of the opportunities they provide for elucidating the molecular basis of transmissible hypovirulence and possible origins of hypovirulence-associated dsRNAs.
136 NAL Call. No.: 381 J824
VMA11, a novel gene that encodes a putative proteolipid, is
indispensable for expression of yeast vacuolar membrane H+-
ATPase activity. Umemoto, N.; Ohya, Y.; Anraku, Y.
Baltimore, Md. : American Society for Biochemistry and
Molecular Biology; 1991 Dec25.
The Journal of biological chemistry v. 266 (36): p.
24526-24532; 1991 Dec25. Includes references.
Language: English
Descriptors: Saccharomyces cerevisiae; Structural genes; Proteolipids; Gene expression; Membranes; Vacuoles; Adenosinetriphosphatase; Amino acid sequences; Nucleotide sequences
Abstract: A gene, VMA11, is indispensable for expression of the vacuolar membrane H+-ATPase activity in the yeast Saccharomyces cerevisiae (Ohya, Y., Umemoto, N., Tanida, I., Ohta, A., Iida, H., and Anraku, Y. (1991) J. Biol. Chem. 266, 13971-13977). The VMA11 gene was isolated from a yeast genomic DNA library by complementation of the vma11 mutation. The nucleotide sequence of the gene predicts a hydrophobic proteolipid of 164 amino acids with a calculated molecular mass of 17,037 daltons. The deduced amino acid sequence shows 56.7% identity, and significant coincidence in amino acid composition with the 16-kDa subunit c (a VMA3 gene product) of the yeast vacuolar membrane H+-ATPase. VMA11 and VMA3 on a multicopy plasmid did not suppress the vma3 and vma11 mutation, respectively, suggesting functional independence of the two gene products. Biochemical detection of the VMA11 gene product was unsuccessful, but vacuoles in the VMA11-disrupted cells were not assembled with either subunit c or subunits a and b of the H+-atpase, resulting in defects of the activity and in vivo vacuolar acidification.
137 NAL Call. No.: 442.8 J828
Yeast cell cycle protein CDC48p shows full-length homology to
the mammalian protein VCP and is a member of a protein family
involved in secretion, peroxisome formation, and gene
expression.
Frohlich, K.U.; Fries, H.W.; Rudiger, M.; Erdmann, R.;
Botstein, D.; Mecke, D. New York, N.Y. : Rockefeller
University Press; 1991 Aug.
The Journal of cell biology v. 114 (3): p. 443-453; 1991 Aug.
Includes references.
Language: English
Descriptors: Saccharomyces cerevisiae; Multigene families; Plant proteins; Cloning; Nucleotide sequences; Amino acid sequences; Restriction mapping; Cell division; Immunocytochemistry; Comparisons; Animal proteins; Mammals; Microsomes
Abstract: Yeast mutants of cell cycle gene cdc48-1 arrest as large budded cells with microtubules spreading aberrantly throughout the cytoplasm from a single spindle plaque. The gene was cloned and disruption proved it to be essential. The CDC48 sequence encodes a protein of 92 kD that has an internal duplication of 200 amino acids and includes a nucleotide binding consensus sequence. Vertebrate VCP has a 70% identity over the entire length of the protein. Yeast Sec18p and mammalian N-ethylmaleimide-sensitive fusion protein, which are involved in intracellular transport, yeast Pas1p, which is essential for peroxisome assembly, and mammalian TBP-1, which influences HIV gene expression, are 40% identical in the duplicated region. Antibodies against CDC48 recognize a yeast protein of apparently 115 kD and a mammalian protein of 100 kD. Both proteins are bound loosely to components of the microsomal fraction as described for Sec18p and Nethylmaleimide -sensitive fusion protein. This similarity suggests that CDC48p participates in a cell cycle function related to that of N-ethylmaleimide-sensitive fusion protein/sec18p in Golgi transport.
Achstetter, T. 44
Alberghina, L. 107
Allen, J. 21
Allen, S.C. 112
Altabella, T. 120
Amore, R. 14
Anderson, J.A. 67
Anraku, Y. 72, 96, 132, 136
Archer, D.B. 111
Arimura, H. 15
Asgeirsdottir, S.A. 126
Atomi, H. 77
Audino, D.C. 7
Auer, B. 83
Aust, S.D. 74
Baetselier, A. de 38
Bandlow, W. 64
Barbetti, M.J. 37
Barnes, C.A. 56
Bartley, G.E. 93
Begdadi-Rais, C. 32
Beier, D. 63
Beier, H. 63, 106
Berberian, G.E. 69
Berka, R.M. 124
Berka, T.R. 21
Bernard, M. 32
Bertheau, Y. 17
Bhairi, S. 10
Bhairi, S.M. 130
Birchmeier, C. 62
Blachly-Dyson, E. 108
Black-Schaefer, L. 89
Blaiseau, P.L. 17
Blomqvist, K. 12
Bodley, J.W. 115
Bominaar, A.A. 117
Bonneaud, N. 16
Bonner, J.J. 134
Borgford, T.J. 42
Botstein, D. 137
Bourbonnais, Y. 75
Bouzayen, M. 79
Brandt, R. 117
Breeden, L. 122
Bron, C. 32
Brown, L.M. 128
Bruenn, J.A. 66
Brygoo, Y. 17
Buloichik, A.A. 99
Bunkers, G.J. 60
Bussink, H.J.D. 46
Buxton, F.P. 46
Cannons, A.C. 49
Cano-Canchola, C. 31
Caput, D. 13
Carmo-Fonseca, M. 41
Carr, C. 6
Carron, C.P. 6
Cauwelaert, F. van 38
Ceccarelli, A. 117
Chamovitz, D. 93
Chiang, C.C. 54, 70
Choi, G.H. 29, 91, 135
Chotai, D. 4
Chrispeels, M.J. 120
Ciriacy, M. 14
Citavicius, D. 84
Clements, J.M. 123
Colicelli, J. 62
Collins, W.J. 37
Colombini, M. 108
Conneely, O.M. 98
Connerton, I.F. 23
Corradin, G. 32
Cox, B. 127
Cregg, J.M. 76
Curtis, P.J. 40
Danoff, A. 75
Davies, R.W. 55
Davio, M. 18
De Zoysa, P.A. 23
Decroupette, I. 118
Deforce, L. 82
Dekker, E.L. 127
Delpech, B. 13
Denis, C.L. 7
Deschenes, R.J. 19
Deutekom, J.C.T. van 131
Dickinson, C.D. 120
Dignard, D. 48
Dimarcq, J.L. 44
Dittrich, H. 94
Dixon, J.E. 19
Dubois, E. 20
Dujon, B. 119
Dumont, X. 13
Durand, R. 95
Duronio, R.J. 6
Dykes, C.W. 4
Eichinger, L. 35
Engelke, D.R. 58
Erdmann, R. 137
Evans, C.F. 58
Farrens, D. 82
Fasel, N. 32
Fassler, J.S. 116
Feaver, W.J. 11
Ferguson, C. 69
Ferrara, P. 13
Fevre, M. 95
Fingar, S.A. 123
Firtel, R.A. 80
Fleig, U.N. 50
Fonzi, W. 31
Forlani, N. 107
Forte, M. 108
Fothergill-Gilmore, L.A. 112
Fraser, J. 133
French, F.S. 4
Fried, M. 101
Friedrich, B. 53
Fries, H.W. 137
Frohlich, K.U. 137
Fujimura, T. 65
Furuya, M. 82
Gaber, R.F. 67
Gaisne, M. 30
Gaymard, F. 16
Germain, D. 48
Giallongo, A. 101
Gillespie, D.J. 37
Gingerich, G. 116
Gleason, W.B. 115
Gold, M.H. 85
Gomi, K. 26
Goodhue, C.T. 123
Gordon, J.I. 6
Gough, K.H. 57
Gralla, E.B. 1
Gray, W. 116
Greaves, P.A. 131
Greenberg, M.L. 105
Grierson, D. 79
Griggs, D.W. 113
Grignon, C. 16
Grison, R. 17
Grivell, L.A. 78
Guan, K. 19
Guillemot, J.C. 13
Haas, M.J. 21
Haastert, P.J.M. van 117
Haber, J.E. 119
Hadwiger, J.A. 80
Hadwiger, L.A. 54, 70
Haguenauer-Tsapis, R. 2
Hahlbrock, K. 104
Haid, A. 64
Halpert, J.R. 68
Hambidge, S.J. 34
Hamilton, A.J. 79
Hanau, R.M. 71
Hara, S. 26
Harkki, A. 92
Hebraud, M. 95
Heinen, Ute 90
Hermann, H. 64
Hickey, D.R. 123
Hinnen, A. 2
Hirschberg, J. 93
Hobden, A.N. 4
Hoffmann, J.A. 44
Hohmann, S. 103
Hohn, T.M. 51
Hollenberg, C.P. 14
Hondel, C.A.M.J.J. van 131
Horaitis, O. 81
Hori, K. 22
Horiuchi, H. 59
Horovitz, D. 54
Hosaka, K. 36, 47
Hosokawa, Y. 123
Huffel, C. van 20
Huprikar, S.S. 67
Hurt, E.C. 41
Icho, T. 65
Iida, H. 96
Iida, N. 49
Irie, M. 59
Ishida, Y. 15
Ito, N. 82
Iwase, T. 114
Jagadish, M.N. 57
Jansen, R.P. 41
Jarvis, E.E. 128
Jayaraman, K. 123
Jeenes, D.J. 111
Johnson, A.L. 28, 122
Johnson, T.M. 73
Johnston, G.C. 56
Johnston, J.R. 23
Johnston, L.H. 28, 122
Johnston, M. 113
Joniau, M. 38
Joos, H.J. 104
Joris, B. 118
Judelson, H.S. 125, 129
Kaiser, P. 83
Kajiwara, S. 22
Kalmar, G.B. 42
Kanamori, J. 109
Karkhoff-Schweizer, R.R. 105
Katayose, Y. 22
Kato, R. 9
Kawabe, H. 15
Kawasaki, H. 132
Kawato, S. 114
Kedzie, K.M. 68
Kelly, B.L. 105
Keppi, E. 44
Kern, H. 41
Kim, C.S. 109
Kinal, H. 66
Kinghorn, J.R. 131
Kirk, N. 87
Kirkegaard, K. 34
Kitamoto, K. 26
Kito, M. 109
Klar, A.J.S. 25
Knowles, J. 12, 92
Kochian, L.V. 67
Kodaki, T. 36
Komel, R. 3
Kondo, J. 77
Korte, A. 5, 102
Kotter, P. 14
Kotula, L. 40
Krabusch, M. 8
Kues, U. 53
Kunz, C. 17
Kuster, C. 14
Kutchan, T.M. 94
Kuyvenhoven, A. 131
Labbe-Bois, R. 30
Lacroute, F. 16
Langner, C.A. 6
Lapeyre, B. 41
LaPorte, D.C. 115
Larson, T.G. 91
Lecocoq, J.P. 44
Lee, J.P. 116
Lee, J.Y. 58
Legoux, R. 13
Legrain, M. 44
Lehtonen, H. 41
Li, J.K.K. 73
Linz, J.E. 33
Livingston, D.M. 115
Lockridge, O. 18
Loison, G. 13
Londesborough, J. 132
London, A. 45
Loper, J.C. 97
Loppes, R. 118
Lottspeich, F. 102
Lowndes, N.F. 28, 122
Lubahn, D.B. 4
Lucas, W.J. 67
Lukins, H.B. 110
Luo, S. 24
Luu, H.A. 133
MacKenzie, D.A. 111
Macreadie, I.G. 81
Madgwick, P.J. 52
Maid, U. 121
Martegani, E. 107
Masaki, A. 15
Massey, V. 18
McAlister-Henn, L. 43
McCourt, J.D. 89
McDonnell, D.P. 98
McKee, E.E. 89
McKown, R.L. 39
McNally, T. 112
Mecke, D. 137
Meskauskas, A. 84
Messenguy, F. 20
Michaelis, U. 5, 102
Michelmore, R.W. 125, 129
Michels, R. 118
Minard, K.I. 43
Minet, M. 16
Minnerly, J.C. 6
Miyazawa, H. 22
Monod, M. 2
Motai, H. 15
Muirhead, H. 112
Mullineaux, P.M. 127
Murakami, K. 15
Murakami, S. 15
Murcott, T.H.L. 112
Nakajima-Shimada, J. 96
Nakamoto, R.K. 61
Nakano, E. 15
Neculcea, J. 133
Nevalainen, K.M.H. 92
Nicolette, C. 62
Nies, D.H. 53
Nikawa, J. 36, 47
Noegel, A.A. 35
Nurse, P. 50
Nuss, D.L. 29, 91, 135
O'Malley, B.W. 98
Oda, K. 77
Oechsner, U. 64
Ohgi, K. 59
Ohkawa, H. 114
Ohlrogge, J.B. 51
Ohta, Y. 114
Ohya, Y. 72, 132, 136
Orlowski, M. 33
Palmgren, M.G. 69
Panaccione, D.G. 71
Pauley, A.M. 6
Payne, M.J. 110
Pearlman, R.E. 11
Pease, E.A. 74
Pecker, I. 93
Peery, R.B. 24
Pel, H.J. 78
Peng, S. 108
Penttila, M. 12
Penttila, M.E. 92
Perentesis, J.P. 115
Perie, F.H. 85
Perrin, A. 119
Peters, D.J.M. 117
Petit, I. 44
Phan, L.D. 115
Philpot, R.M. 68
Pines, O. 45
Piper, P.W. 87
Plessis, A. 119
Poletti, A. 98
Pouwels, P.H. 131
Poyton, R.O. 89
Pratt, K.A. 52
Punt, P.J. 131
Purvis, I.J. 4
Qiu, H. 19
Racher, K.I. 42
Ramond, P. 13
Rao, R. 61
Reed, S.I. 27
Reichhart, J.M. 44
Rensing, C. 53
Reymond, C.D. 32
Reymond, P. 95
Riggs, M. 62
Roberts, I.N. 111
Rodel, G. 5, 102
Rodgers, L. 62
Rogers, P.V. 24
Rozman, D. 3
Rudiger, M. 137
Ruiz-Herrera, J. 31
Russell, P.J. 34
Rutherford, C.L. 24
Ruusala, T. 86
Saito, K. 18
Saito, T. 22
Sakaki, T. 114
Salmon, J.M. 16
Sato, T. 109
Savin, K.W. 81
Schaap, P. 117
Schleicher, M. 35
Schrader, W.T. 98
Schweiger, M. 83
Schweizer, E. 110
Scolnik, P.A. 93
Selmin, O. 24
Sentenac, H. 16
Serrano, R. 69
Shah, J. 123
Shapira, R. 29, 135
Sherman, F. 123
Shewry, P.R. 52
Shields, D. 75
Shire, D. 13
Shishido, K. 22
Shukla, D.D. 57
Sierkstra, L.N. 100
Silar, P. 1
Silve, S. 2
Singer, R.A. 56
Slayman, C.W. 61
Snaar-Jagalska, B.E. 117
Solomonson, L.P. 49
Song, P.S. 82
Sosa, L. 31
Springer, J. 126
Stahl, U. 53
Staples, R.C. 10, 130
Steffan, J.S. 43
Steinmuller, R. 121
Storms, R.K. 133
Strathmann, M. 80
Sucic, J.F. 24
Sudberry, P.E. 88
Suihko, M.L. 12
Sun, G.H. 72
Suzuki, K. 132
Sypherd, P. 31
Szweykowska-Kulinska, Z. 106
Tada, S. 26
Takagi, M. 59
Takahashi, K. 26
Tamura, G. 26
Tanaka, A. 77
Tao, J. 66
Tatsumi, H. 15
Teeri, T.T. 92
Teranishi, Y. 77
Tessier, D. 48
Thiele, D.J. 1, 18
Thomas, D.Y. 48, 75, 133
Thon, G. 25
Tien, M. 74
Togni, G. 2
Tollervey, D. 41
Tomizawa, K.I. 82
Tschopp, J.F. 76
Tsuji, F.I. 96
Tsuji, R.F. 15
Tsukagoshi, Y. 47
Tsunasawa, S. 123
Tulloch, P.A. 57
Turi, T.G. 97
Tzagoloff, A. 78
Ueda, M. 77
Umemoto, N. 136
Utsumi, S. 109
Valentine, J.S. 1
Veale, R.A. 88
Verbakel, J.M.A. 100
Verdiere, J. 30
Verkuylen, A.J. 81
Verrips, C.T. 100
Viaene, A. 38
Viitanen, P.V. 93
Villalba, J.M. 69
Visser, J. 46
Voluevich, E.A. 99
Vries, O.M.H. de 126
Vvolckaert, G. 38
Ward, C.W. 57
Warren, G.J. 39
Watanabe, H. 59
Watson, D.C. 23
Weigel, N.L. 98
Wessels, J.G.H. 126
Whiteway, M. 48
Whittaker, L.A. 57
Wickner, R.B. 65
Widner, W.R. 65
Wiegand, R.C. 6
Wigler, M. 62
Wilkie, T.M. 80
Williams, J.G. 117
Wilson, E.M. 4
Wittenberg, C. 27
Wolf, K. 8
Woolston, C.J. 127
Xue, Y. 127
Xuei, X. 10
Yabusaki, Y. 114
Yamashita, S. 36, 47
Yamazoe, Y. 9
Yang, Z.F. 71
Yano, K. 59
Yasumori, T. 9
Yin, Y. 24
Yoder, O.C. 10
Yon, J. 101
Yu, G. 116
Zetsche, K. 121
Zimmer, M. 8
Zollner, A. 64
Acetylglutamic acid 20
Acid phosphatase 2
Actin 35, 91
Adenosinetriphosphatase 61, 69, 136
Adp 83
Aegilops ovata 99
Aegilops squarrosa 99
Air temperature 125
Alcohol dehydrogenase 7
Alcohol oxidoreductases 88
Alkanes 77
Alleles 50, 103
Alpha-amylase 26
Alpha-galactosidase 100
Alpha-lactalbumin 38
Amino acid metabolism 107
Amino acid sequences 6, 8, 10, 13, 14, 15, 16, 17, 18, 19,
20, 21, 22, 23, 24, 38, 41, 44, 45, 46, 48, 49, 50, 59, 62,
67, 70, 79, 80, 82, 84, 91, 93, 94, 101, 102, 103, 104, 109,
115, 118, 121, 123, 132, 136, 137
Anabolism 20
Anaerobes 95
Anaerobic conditions 30
Androgens 4
Androstenedione 3
Animal proteins 137
Antibacterial properties 44
Antigens 32
Arabidopsis thaliana 16, 63, 67, 69, 106
Arginine 20
Argininosuccinate lyase 118
Asexual reproduction 71
Aspergillus 46
Aspergillus flavus 13
Aspergillus nidulans 3, 55, 131
Aspergillus niger 46, 55, 111, 124
Aspergillus oryzae 15, 26
Atp 110
Autographa californica 74
Bacterial proteins 39
Beers 12
Beta-fructofuranosidase 100, 120
Beta-galactosidase 131
Beta-glucuronidase 26, 60, 129, 130, 131
Binding proteins 80, 117, 134
Binding site 1, 15, 35, 49, 64, 83
Biological control 135
Biological development 22
Biosynthesis 36, 51, 79, 82, 92, 94, 97, 107
Blight 135
Bremia lactucae 125, 129
Brewing industry 12
C-amp 7, 22, 24, 117
Calcium ions 96
Calmodulin 72, 132
Candida albicans 6
Candida tropicalis 77
Carbohydrate metabolism 26
Carboxy-lyases 12
Carotenoids 93
Castanea 91
Castanea sativa 135
Cauliflower mosaic caulimovirus 120
Cell differentiation 10, 24
Cell division 27, 50, 72, 83, 137
Cell growth 96
Cell ultrastructure 41
Cell wall components 2
Cellulolytic microorganisms 92
Cellulose 92
Cellulose digestion 92
Characterization 38, 74, 112
Chemical reactions 83
Chitinase 17
Chlorella ellipsoidea 128
Chlorella vulgaris 49
Chloroplast genetics 121
Choline 36, 105
Chromatin 11
Chromosomes 116
Chymosin 124
Clones 20, 45, 49, 104
Cloning 13, 14, 15, 16, 18, 19, 21, 22, 24, 32, 42, 47, 48,
62, 66, 67, 75, 84, 93, 94, 95, 101, 115, 123, 132, 137
Coat proteins 57
Cochliobolus lunatus 3
Coenzyme a 6
Cold resistance 39
Colletotrichum graminicola 71
Comparisons 91, 101, 137
Conidia 71
Controlling elements 25, 30, 64, 98
Copper 1
Crop damage 37
Crop growth stage 37, 99
Cryoprotectants 39
Cryphonectria parasitica 29, 91, 135
Cultivars 37, 54
Cyamopsis tetragonoloba 100
Cysteine 70
Cytochrome b 5
Cytochrome c 64, 123
Cytochrome 9, 30, 97, 114
Cytochrome-c oxidase 78
Cytochromes 68
Cytoplasmic inclusions 61
Cytoplasmic inheritance 8, 99
Deficiency 30
Deletions 119
Deuteromycotina 17
Developmental stages 22, 31
Diacetyl 12
Dictyostelium 24, 32, 80, 117
Diphosphatidylglycerols 105
Disease resistance 37, 51, 54, 70, 125
Dna 28, 31, 59, 62, 66, 70, 83, 93, 104, 121, 122
Dna binding proteins 7, 30, 64, 116
Dna hybridization 95
Dna libraries 95
Dna modification 119
Dna repair 119
Dna sequencing 11
Dominance 54
Dominant lethals 50
Drug resistance 66
Endomycetales 8, 25, 50, 86, 100
Endoplasmic reticulum 2
Enterobacter aerogenes 12
Environmental factors 125
Enzyme activity 1, 2, 7, 12, 14, 15, 20, 21, 24, 42, 43, 45,
47, 48, 50, 51, 60, 61, 74, 77, 79, 83, 85, 94, 103, 105, 111,
118, 119, 123
Enzyme precursors 15
Enzymes 97
Ergosterol 97
Escherichia coli 3, 21, 26, 43, 47, 57, 60
Eschscholzia californica 94
Ethylene 79
Extracts 63
Fermentation 76, 87
Food industry 124
Food research 109
Fowls 98
Freezing 39
Fruiting 126
Fungal antagonists 17
Fungal diseases 29, 94, 99
Fungal morphology 126
Fungal protein 124
Fungi 35, 55
Fusarium oxysporum 17
Fusarium oxysporum f.sp. pisi 54
Fusarium solani f.sp. phaseoli 70
Fusarium solani f.sp. pisi 70
Fusarium sporotrichioides 51
Gene conversion 119
Gene expression 1, 3, 4, 5, 6, 7, 9, 10, 11, 12, 13, 14, 15,
16, 17, 18, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31,
32, 33, 35, 36, 38, 39, 40, 42, 43, 44, 45, 46, 47, 48, 51,
52, 53, 54, 55, 56, 58, 59, 60, 62, 63, 64, 68, 70, 71, 72,
74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 86, 87, 88, 89, 91,
92, 93, 94, 95, 96, 97, 98, 100, 102, 103, 104, 105, 107, 109,
110, 112, 113, 114, 115, 116, 117, 118, 119, 120, 121, 122,
123, 124, 125, 126, 127, 128, 129, 130, 131, 132, 133, 136
Gene location 25, 116
Gene mapping 25, 80, 93, 116, 118, 128
Gene transfer 21, 55, 83, 88, 100
Genes 4, 5, 7, 8, 10, 11, 12, 14, 15, 17, 20, 21, 23, 25, 28,
30, 31, 32, 36, 45, 48, 54, 56, 70, 78, 83, 91, 98, 101, 102,
116, 121, 125, 126, 131, 133
Genetic analysis 60, 70, 80, 91
Genetic change 86
Genetic code 20, 35, 40, 41, 46, 47, 58, 62, 77, 80, 84, 92,
103, 109, 118
Genetic control 29
Genetic engineering 26, 40, 73, 98, 103, 109, 124
Genetic markers 60
Genetic regulation 7, 11, 17, 25, 27, 30, 36, 64, 78, 92, 97,
103, 105, 106, 107, 110, 113, 116, 117, 127, 130, 133
Genetic resistance 54, 99
Genetic transformation 3, 9, 12, 14, 15, 17, 26, 38, 39, 40,
43, 53, 55, 60, 66, 68, 76, 79, 83, 87, 88, 100, 102, 111,
120, 128, 129, 130
Genomes 8, 104
Genotype environment interaction 125
Genotypes 125
Gliadin 52
Glucose 64, 113
Glutamate dehydrogenase 23
Glyceraldehyde-3-phosphate dehydrogenase 91
Glycine max 93, 109
Glycine soja 93
Glycogen phosphorylase 24
Growth 120
Guanine 80
Hansenula 88
Heat shock 56, 62, 87, 121, 133, 134
Hela cells 63
Heme 30, 49, 64
Hordeum vulgare 60
Hormone receptors 98
Horses 123
Host parasite relationships 54, 70
Host specificity 125
Hydrolases 95
Hygromycin b 66
Hyperparasitism 17
Hyphae 10, 126
Hypovirulence 29, 91, 135
Immunocytochemistry 109, 137
Immunoglobulin structural genes 40
In vitro 63
Induced mutations 25, 48, 50, 78
Induction 87, 95
Industrial microbiology 124
Infections 10
Infectivity 130
Inhibition 120
Inhibitors 113
Injuries 104
Inositol phosphates 117
Introns 10, 22, 91, 106, 119
Ion transport 67
Ions 53
Isocitrate lyase 77
Isoenzymes 9, 73, 74
Isolation 6, 132
Isoleucine 42
Kabatiella caulivora 37
Kinases 20
Kinetics 33, 42, 105
Klebsiella 12
Laccase 85, 91
Lactuca sativa 125
Lanosterol 97
Lentinula edodes 22
Ligases 42, 51, 105, 110
Light 120
Lignin 73, 85
Ligninolytic microorganisms 73
Lines 54
Literature reviews 27, 55, 88, 92
Loci 25, 86
Luciferase 128
Luminescence 96
Lycopersicon esculentum 79, 120
Lysozyme 111
Malate dehydrogenase 43
Mammals 68, 75, 137
Man 4, 9, 83
Manganese 74, 85
Marker genes 66
Mating 25, 86, 116
Meiosis 25
Membranes 105, 108, 136
Mercury 53
Messenger RNA 5, 10, 17, 25, 26, 33, 34, 63, 70, 78, 91,
101, 104, 116, 123, 133
Metabolism 12
Metal ions 1
Methanol 76, 87, 88
Methionine 107
Methyl methanesulfonate 133
Methylation 31
Mice 40, 101
Microbial degradation 85
Microbial proteins 77
Microsomes 114, 137
Mildews 99
Mineralization 85
Mitochondria 5, 20, 43, 89, 102, 105, 110
Mitochondrial DNA 8, 78, 119
Mitochondrial genetics 8, 78
Molds 117
Molecular biology 92
Molecular conformation 63, 108
Molecular genetics 79, 94
Molecular mapping 127
Molecular weight 70
Mucor 31, 33
Multigene families 137
Multiple genes 63
Mutagenesis 112
Mutagens 133
Mutants 11, 20, 25, 35, 48, 50, 67, 78, 82, 92, 103, 108,
110, 126 Mutations 8, 36, 63, 108, 113, 116, 126
Mutator genes 8
Mycelium 22
Mycotoxins 48, 66, 84
Myo-inositol 36, 105
Myristic acid 6
Nad 14
Nadh 14
Nadp 23
Nadph 18
Nadph-cytochrome-c2 reductase 114
Neocallimastix 95
Neurospora crassa 91
Nicotiana rustica 106
Nicotiana tabacum 51
Nitrate reductase 49, 131
Nitrite reductase 131
Nuclear polyhedrosis viruses 74
Nucleases 45, 119
Nuclei 11, 78, 119
Nucleocytoplasmic interaction 78, 99
Nucleoproteins 83
Nucleotide sequences 6, 8, 10, 13, 14, 15, 16, 17, 18, 19,
20, 21, 22, 23, 24, 29, 38, 39, 41, 44, 46, 48, 49, 50, 58,
59, 62, 63, 64, 66, 67, 70, 79, 80, 84, 91, 93, 94, 95, 100,
101, 103, 104, 109, 115, 116, 118, 121, 123, 127, 132, 134,
135, 136, 137
Oncogenes 22, 62
Organelles 77
Ornithine carbamoyltransferase 20
Outbreaks 37
Oxidoreductases 13, 14, 18, 20, 30, 53, 74, 79, 85, 94, 100
Oxygen 30, 64
Pathogenesis 10
Pathogenesis-related proteins 70
Pathogenicity 37
Pentosyltransferases 83
Peptides 44
Peroxidases 73, 85
Ph 38
Phanerochaete chrysosporium 73, 74
Phaseolus vulgaris 130
Phenotypes 8, 84, 120, 126
Phenylalanine ammonia-lyase 104
Phormia terraenovae 44
Phospholipase a2 111
Phospholipids 117
Phosphoproteins 50
Phosphoric monoester hydrolases 19
Phosphorylation 19
Photosynthates 120
Phytoalexins 94
Phytochrome 82
Phytophthora infestans 104, 129
Phytotoxicity 48
Pichia 88
Pichia stipitis 14
Pigeons 123
Pisum sativum 54, 70
Plant breeding 99
Plant development 99
Plant disease control 135
Plant extracts 63
Plant pathogenic fungi 91, 99, 129
Plant pathogens 60
Plant proteins 2, 5, 35, 70, 84, 96, 98, 102, 121, 133, 137
Plant viruses 135
Plasma membranes 5, 61, 69
Plasmids 3, 47, 55, 66, 68, 69, 75, 86, 88, 100, 134
Plasmodium falciparum 32
Plastids 121
Polygalacturonase 46
Polymerase chain reaction 19, 80, 127
Polypeptides 10
Polyporus 85
Potassium 16, 67
Potyvirus group 57
Precursors 63, 75
Progesterone 3, 98
Promoters 4, 20, 26, 32, 58, 64, 77, 87, 88, 98, 103, 113
Protein composition 35
Protein kinase 7, 50, 132
Protein quality 109
Protein secretion 38, 45, 111
Protein synthesis 2, 33, 35, 40, 41, 89
Proteinases 15, 48, 111
Proteins 32, 55, 65, 88, 101, 108, 122, 126
Proteolipids 136
Proteolysis 29, 89, 102, 111
Protoplasts 128
Pseudocercosporella herpotrichoides 60
Pseudogenes 63
Purification 24, 74, 102, 112
Pyruvate decarboxylase 103
Pyruvate kinase 112
Races 54
Rats 43, 114
Receptors 4
Recombinant DNA 4, 17, 18, 26, 32, 42, 55, 56, 69, 81, 96,
98, 131, 134
Recombinant vaccines 32
Recombination 25, 38, 74, 77, 82, 112, 119
Refeeding 133
Regulation 5, 24
Replication 65
Reporter genes 26, 87, 100, 128, 129
Resistance 48, 53, 84
Restriction fragment length polymorphism 93
Restriction mapping 17, 46, 63, 64, 103, 116, 121, 137
Rhizopus 21, 59
Rhodophyta 121
Ribonucleases 58, 59
Ribose 83
Ribosomes 101
Rna 48, 65, 84, 95, 127, 135
Rna editing 63, 78
Rna polymerase 58
Rumen fungi 95
Saccharomyces 28
Saccharomyces cerevisiae 1, 2, 4, 5, 7, 9, 11, 12, 14, 15,
16, 19, 20, 23, 27, 30, 31, 34, 36, 38, 39, 40, 42, 43, 44,
45, 47, 48, 52, 53, 56, 57, 58, 59, 61, 62, 64, 65, 67, 68,
72, 75, 77, 78, 79, 81, 82, 83, 84, 87, 89, 94, 96, 97, 98,
100, 101, 102, 103, 107, 108, 109, 110, 112, 113, 114, 115,
116, 118, 119, 123, 132, 133, 134, 136, 137
Saccharomyces uvarum 18
Schizophyllum commune 126
Screening 37, 95
Secale cereale 60
Secretory granules 61
Segregation 8
Semidominance 25
Serine 50, 63
Sesquiterpenes 51
Sex pheromones 96
Solanum tuberosum 104
Somatostatin 75
Soy protein 109
Species differences 91
Spectral analysis 49, 114
Sporangia 33
Spore germination 33
Spores 33
Sporulation 99
Staphylococcus aureus 45
Starch 26
Starvation 133
Steroid metabolism 3
Strain differences 91
Strains 12, 20, 58, 62, 91, 113, 117
Stress response 70, 104
Structural genes 6, 13, 24, 55, 64, 88, 103, 104, 115, 132,
136 Structure activity relationships 72
Substitution lines 99
Substrates 76, 88
Sugars 26
Superoxide dismutase 1
Suppression 106, 126
Survival 39, 87
Susceptibility 37
Symptoms 37, 120
Synergism 113
Synthesis 28
Synthetic genes 38, 123
Targeted mutagenesis 48
Temperature 105, 110
Testosterone 3
Thigmotropism 10
Thunnus 123
Tolerance 87
Toxicity 66
Transcription 1, 7, 22, 25, 27, 30, 31, 56, 58, 63, 64, 78,
86, 95, 98, 101, 110, 113, 116, 121, 122, 127, 133
Transfer RNA 42, 63, 106
Transferases 6, 47
Transformation 69
Transgenics 15, 51, 120
Translation 5, 8, 20, 29, 89, 95, 121
Translocation 34, 120
Transport processes 61
Transposable elements 11, 86, 116
Triacylglycerol lipase 21
Trichoderma 92
Trifolium subterraneum 37
Triticum aestivum 60, 99
Triticum spelta 99
Tubulin 91
Tyrosine 19
Uric acid 13
Uromyces appendiculatus 10, 130
Ustilago zeae 66
Vacuoles 136
Varietal susceptibility 54
Vectors 3, 4, 9, 32, 34, 43, 55, 59, 66, 81, 88, 100, 114,
123, 129, 131, 134
Virulence 125
Virus-like particles 66
Western australia 37
Wheat 52
Wheat dwarf geminivirus 127
Wheat germ 63
Xylose 14
Yeast extracts 63
Yeasts 1, 20, 23, 41, 62, 67, 69, 76, 82, 105, 113, 118, 122
Zea mays 71
Zinc 1, 15