TITLE: Gene Expression in Algae and Fungi Including YeastPUBLICATION DATE:January, 1993ENTRY DATE: March, 1994EXPIRATION DATE: NoneUPDATE: As neededCONTACT: Biotechnology Information Center(biotech@nalusda.gov)         National Agricultural LibraryDOCUMENT TYPE: TextDOCUMENT SIZE: 232.9k, approx. 129 pp.
                                             ISSN:  1052-5378United States Department of Agriculture      National Agricultural Library10301 Baltimore Blvd.Beltsville, Maryland  20705-2351
Gene Expression in Algae and Fungi Including YeastJanuary 1991 - November 1992
QB 93-10
137 citations from AgricolaJanet Saunders and Robert WarmbrodtBiotechnology Information Center
January 1993 National Agricultural Library Cataloging Record:
Saunders, Janet  Gene expression in algae and fungi, including yeast.  (Quick bibliography series ; 93-10) -  Gene expression--Bibliography. 2. Algae--Genetic aspects--Bibliography. 3. Fungi--Genetic aspects--Bibliography. 4.Yeast--Genetic aspects--Bibliography. I. Warmbrodt, Robert D.II. Title.aZ5071.N3 no.93-10 AGRICOLA  SEARCH STRATEGY
Set   Items     DescriptionS1    5992      EXPRESS?/TIS2   99924      ALGA? OR DEUTEROMYCOTINA OR FUNG? OR MOLD? OR                MOULD? OR YEAST?S3     705      S1 AND S3S4     166      S3 AND PY=1991:1992
Gene Expression in Algae and Fungi, Including Yeast
1                                   NAL Call. No.: 500 N21PACE1, a copper-dependent transcription factor, activatesexpression of the yeast copper, zinc superoxide dismutasegene.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 UnitedStates 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 geneproduct) (superoxide:superoxide oxidoreductase, EC 1.15.1.1)is a copper-containing enzyme that functions to prevent oxygentoxicity. In the yeast Saccharomyces cerevisiae, copper levelsexert some control over the level of SOD1 expression. We showthat the ACE1 transcriptional activator protein, which isresponsible for the induction of yeast metallothionein (CUP1)in response to copper, also controls the SOD1 response tocopper. A single binding site for ACE1 is present in the SOD1promoter region, as demonstrated by DNase I protection andmethylation interference experiments, and is highly homologousto a high-affinity ACE1 binding site in the CUP1 promoter. Thefunctional importance of this DNA-protein interaction isdemonstrated by the facts that (i) copper induction of SOD1mRNA does not occur in a strain lacking ACE1 and (ii) it doesnot occur in a strain containing a genetically engineered SOD1promoter that lacks a functional ACE1 binding site.
2                                  NAL Call. No.: QH573.N37The acid phosphatase of Saccharomyces cerevisiae: a model tostudy 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 immuneresponse / edited by J.P. Latge and D. Boucias. Proceedings ofthe 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.C8Altered expression of the steroid bioconverting pathway in pAN7-1 transformants of Cochliobolus lunatus.Rozman, D.; Komel, R.Berlin, W. Ger. : Springer International; 1991.Current genetics v. 20 (5): p. 385-389; 1991.  Includesreferences.
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 convertsprogesterone mainly to its 11 beta-hydroxy derivative. C.lunatus transformed with the plasmid pAN 7-1, which containsthe 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 progesteronethrough the reaction scheme progesterone leads to 20 betahydroxy-progesterone leads to delta 4-androstene-3,17-dioneleads to testolactone + testosterone. The main part of thismetabolic pathway is not expressed in the non-transformedstrain. It was determined that the site-specific integrationof the plasmid into the genome directly influences theexpression of genes involved in the bioconversion of steroids.
4                                 NAL Call. No.: QH442.A1G4An androgen-inducible expression system for Saccharomycescerevisiae. 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; Geneexpression; Androgens; Receptors; Genes; Man; Promoters;Recombinant  DNA
Abstract:  A novel controllable expression system forSaccharomyces cerevisiae has been developed. Expression of thegene encoding the human androgen receptor, from a strong yeastpromoter, results in transactivation of a hybrid promotercarrying androgen-responsive sequences such that a target genemay be expressed in an androgen-dependent manner. By selectionof an appropriate combination of androgen receptor level,target-gene copy number and concentration of the androgenicligand, dihydrotestosterone, the expression level can be setwithin a 1400-fold range with no detectable effect on normalcell growth.
5                                  NAL Call. No.: 442.8 Z34Association of cytochrome b translational activator proteinswith the mitochondrial membrane: implications for cytochrome bexpression 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; Geneexpression; Cytochrome b; Messenger  RNA; Regulation; Genes;Plant proteins; Mitochondria; Plasma membranes
Abstract:  The products of the nuclear genes CBS1 and CBS2 areboth required for translational activation of mitochondrialapocytochrome b in yeast. We report the intramitochondriallocalization of both proteins by use of specific antisera.Based on its solubilization properties the CBS1 protein ispresumed to be a component of the mitochondrial membrane; thedetergent concentrations needed to release CBS1 frommitochondria are almost the same as for cytochrome c1. Incontrast, CBS2 behaves like a soluble protein, with somecharacteristics of a membrane-associated protein. A model ispresented for translational activation of cytochrome b, whichmight also be applicable to translational regulation of othermitochondrial genes.
6                                   NAL Call. No.: 381 J824The Candida albicans myristoyl-CoA:protein Nmyristoyltransferase gene. Isolation and expression inSaccharomyces 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 andMolecular 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 andantifungal therapy. Candida albicans is a dimorphic, asexualyeast that is a major cause of systemic fungal infections inimmunosuppressed humans. metabolic labeling studies indicatethat C. albicans synthesizes one principal 20-kDa N-myristoylprotein. The single copy C. albicans NMT gene (ca-NMT1) wasisolated and encodes a 451-amino acid protein that has 55%identity with Saccharomyces cerevisiae NMT. C. albicans NMT1is able to complement the lethal phenotype of S. cerevisiaenmt1 null mutants by directing efficient acylation of theapproximately 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 studiesof purified E. coli-derived S. cerevisiae and C. albicans NMTsrevealed species-specific differences in the kineticproperties of synthetic octapeptide substrates derived fromknown N-myristoylproteins. Together these data indicate thatC. albicans and S. cerevisiae NMTs have similar yet distinctsubstrate specificities which may be of therapeuticsignificance.
7                                  NAL Call. No.: 442.8 Z34The CCR1 (SNF1) and SCH9 protein kinases act independently ofcAMP-dependent protein kinase and the transcriptionalactivator 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; Geneticregulation; Protein kinase; C-amp; Dna binding proteins;Enzyme activity
Abstract:  cAMP-dependent protein kinase (cAPK) is implicatedin the inactivation of the yeast transcriptional activatorADR1, which regulates glucose-repressible ADH2 geneexpression. The interdependence of cAPK, SCH9 (a proteinkinase that when overexpressed can functionally substitute forcAPK), and the CCR1 (SNF1) protein kinase that is required forADH2 expression was studied. SCH9 was found to be required forADH2 expression in contrast to the inhibitory role played bycAPK. CCR1 and SCH9 were observed to affect ADH2 expressionindependently of both ADR1 and cAPK. In contrast, cAPK wasshown to exert its effects on ADH2 solely through ADR1. Theseresults indicate that the SCH9 and CCR1 protein kinases arecomponents of regulatory pathways separate from that utilizedby cAPK to control ADR1 activity and ADH2 expression.
8                                   NAL Call. No.: QH426.C8Characterization of a novel open reading frame, urf a, in themitochondrial genome of fission yeast: Correlation of urf amutations with a mitochondrial mutator phenotype and apossible 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.  Includesreferences.
Language:  English
Descriptors: Endomycetales; Mitochondrial  DNA; Mitochondrialgenetics; 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) anopen reading frame of 227 amino acids has been identifiedwhich is unique among known mitochondrial genomes and whichhas been termed urf a (Lang et al. 1983; Kornrumpf et al.1984). It uses the "mitochondrial" genetic code, i.e., itcontains a TGA codon, whereas all other protein-encodinggenes, and all but one intronic open reading frame, use the"standard" genetic code (UGG for tryptophan). A previous paperhas demonstrated that "mutator" strains show an increasedformation of mitochondrial drug-resistant and respirationdeficient mutants (including deletions). In this paper we showthat the mutator activity is correlated with mutations in urf-  A detailed analysis of one urf a mutant is presented(ana(r)-6), where the deletion of an A residue leads to aframeshift mutation and consequently to premature terminationof the putative protein. The phenotype of colonies originatingfrom a single mutant clone varies from no growth up to fullgrowth on non-fermentable substrate. This phenomenon ofphenotypic segregation can be explained by the ability of thecell to perform translational frameshifting. A detailedanalysis of the DNA sequence and the putative urf a proteinwill be presented and a possible function of the protein willbe discussed.  9                                  NAL Call. No.: QP601.M49Characterization of human P450IIC isozymes by using yeastexpression 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 seriesanalytic: Cytochrome P450 / edited by M.R. Waterman and E.F.Johnson.  Includes references.
Language:  English
Descriptors: Cytochrome p-450; Isoenzymes; Vectors; Geneexpression; Genetic transformation; Man; Saccharomycescerevisiae
10                                NAL Call. No.: QH442.A1G4Characterization of INF56, a gene expressed during infectionstructure 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; Geneexpression; Messenger  RNA; Hyphae; Cell differentiation;Infections; Pathogenesis; Thigmotropism
Abstract:  The bean rust fungus, Uromyces appetidiculatus,undergoes thigmotropic differentiation to produce infectionstructures. Six differentiation-specific genes have beenisolated and one, INF24, has been characterized [Bhairi etal., Gene 81 (1989) 237-243]. Here, we report the structure ofa second gene, IVF56, which was subcloned on a 2.6-kb fragmentand sequenced. The location of the 1.0-kb INF56 transcript wasdetermined by S1 nuclease protection and primer extension. ATATA box was found 38 bp upstream and a CAAT box 130 bpupstream from the major transcription start point (tsp). Thegene contains two open reading frames: ORF2 is nested withinORF1; they share a 67-bp intron. ORF1 encodes a 14.1-kDapolypeptide which has an amino acid sequence rich in Gly, Proand Ser. It has sequence similarity to a functional domain(V2) of mammalian cytokeratin type II. ORF2 encodes a 10.1-kDapolypeptide which is rich in Pro. It shares similarity withthe cell-surface recognition region of chicken fibronectin.Hybrid selection and in vitro translation of the INF56 mRNAyielded two polypeptides of 15.5 and 23 kDa, as estimated bysodium dodecyl sulfate-polyacrylamide-gel electrophoresis.INF56 is constitutively expressed at a low level, but theabundance of its steady-state transcript is upshifted 4.5 hafter spore hydration during the period that infectionstructures are formed.
11                                NAL Call. No.: QP501.B642Chromatin structure in a region of a yeast transposableelement regulating adjacent gene expression.Feaver, W.J.; Pearlman, R.E.Ottawa : National Research Council of Canada; 1991 May.Biochemistry and cell biology; Biochimie et biologiecellulaire v. 69 (5/6): p. 392-398; 1991 May.  Includesreferences.
Language:  English
Descriptors: Saccharomyces cerevisiae; Mutants; Nuclei;Chromatin; Genes; Transposable elements; Gene expression;Genetic regulation; Dna sequencing
12                                 NAL Call. No.: 448.3 AP5Chromosomal 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; 1991Oct. 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-acetolactatedecarboxylase, isolated from Klebsiella terrigena orEnterobacter aerogenes, was expressed in brewer's yeast. Thegenes were expressed under either the yeastphosphoglycerokinase (PGKI) or the alcohol dehydrogenase(ADHI) promoter and were integrated by gene replacement byusing cotransformation into the PGKI or ADHI locus,respectively, of a brewer's yeast. The expression level of thealpha-acetolactate decarboxylase gene of the PGK1 integrantstrains was higher than that of the ADH1 integrants. Underpilot-scale brewing conditions, the alpha-acetolactatedecarboxylase activity of the PGK1 integrant strains wassufficient to reduce the formation of diacetyl below the tastethreshold value, and no lagering was needed. The brewingproperties or the recombinant yeast strains were otherwiseunaltered, and the quality (most importantly, the flavor) ofthe trial beers produced was as good as that of the controlbeer.
13                                  NAL Call. No.: 381 J824Cloning and expression in Escherichia coli of the geneencoding 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 andMolecular 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; Nucleotidesequences; Amino acid sequences
Abstract:  Amino acid sequencing of peptides obtained afterproteolytic hydrolysis of Aspergillus flavus urate oxidase(uricase) permitted the design of oligodeoxynucleotide probesthat were used to obtain 1.2- and 5-kilobase pair DNAfragments from A. flavus cDNA and genomic libraries,respectively. The cDNA fragment contained the entire codingregion for uricase, and comparison with the genomic fragmentrevealed the presence of two short introns in the codingregion of the gene. A. flavus uricase has around 40% overallidentity with uricases from higher organisms but with manyconserved amino acids. Hitherto highly conserved consensuspatterns found in other uricases were found to be modified inthe A. flavus enzyme, notably the sequence Val-Leu-Lys-ThrThr-Gln-Ser near position 150, which in the filamentous fungusis uniquely modified to Val-Leu-Lys-Ser-Thr-Asn-Ser. Silentmutations were introduced by cassette mutagenesis near the 5'-extremity of the coding sequence in order to conform withEscherichia coli codon usage, and the uricase was expressed inthe E. coli cytoplasm in a completely soluble, biologicallyactive form.
14                                NAL Call. No.: QH442.A1G4Cloning and expression in Saccharomyces cerevisiae of theNAD(P)H-dependent xylose reductase-encoding gene (XYL1) fromthe 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; Nucleotidesequences; Amino acid sequences; Genetic transformation; Geneexpression; Enzyme activity
Abstract:  The XYL1 gene of the yeast Pichia stipitis has beenisolated from a genomic library using a specific cDNA probe,and its nucleotide (nt) sequence has been determined. In the5' noncoding region of the P. stipitis XYL1 gene a TATAAAelement (known to be necessary for transcription initiation inmost yeast genes) is found at nt -81, and two CCAATrecognition motifs (often referred to as the CCAAT box) arepresent at nt -146 and -106. The XYL1 encodes a polypeptide of35927 Da that constitutes a NADH/NADPH-dependent xylosereductase (XR). The enzyme is part of the xylose-xylulosepathway that is absent or only weakly expressed inSaccharomyces cerevisiae. Extensive homology is found to the Nterminus of the XR of Pachysolen tannophilus and Candidashehatae. None of the known cofactor binding domains found inmany NAD-dependent dehydrogenases are present in the protein.Transformants of S. cerevisiae containing XYL1 of P. stipitissynthesize an active XR. Fusion of XYL1 with the prokaryotictac promoter leads to a gene that can be expressed in S.cerevisiae and Escherichia coli.
15                                 NAL Call. No.: 442.8 Z34Cloning and expression in yeast of a cDNA clone encodingAspergillus oryzae neutral protease II, a uniquemetalloprotease.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 acidsequences; Genetic transformation; Transgenics; Geneexpression; Enzyme precursors; Zinc; Binding site; Enzymeactivity
Abstract:  The neutral protease II (NpII) from Aspergillusoryzae is a zinc-containing metalloprotease with some uniqueproperties. To elucidate its structure, we isolated a fulllength cDNA clone for NpII. Sequence analysis reveals thatNpII has a prepro region consisting of 175 amino acidspreceding the mature region, which consists of 177 aminoacids. As compared with other microbial metalloproteases, NpIIis found to be unique in that it shares only a limitedhomology with them around two zinc ligand His residues andthat the positions of the other zinc ligand (Glu) and theactive site (His) cannot be established by homology. When aplasmid designed to express the prepro NpII cDNA wasintroduced into Saccharomyces cerevisiae and the transformantwas cultured in YPD medium (2% glucose, 2% polypeptone, 1%yeast extract), it secreted a proNpII. However, in a cultureof the same medium containing 0.2 mM ZnCl2, it secreted amature NpII with a specific activity and N-terminus identicalto those of native NpII. This observation suggests that eitheran autoproteolytic activity or a yeast protease effected theprocessing.
16                                  NAL Call. No.: 470 SCI2Cloning and expression in yeast of a plant potassium iontransport system. Sentenac, H.; Bonneaud, N.; Minet, M.;Lacroute, F.; Salmon, J.M.; Gaymard, F.; Grignon, C.Washington, D.C. : American Association for the Advancement ofScience; 1992 May01.Science v. 256 (5057): p. 663-665; 1992 May01.  Includesreferences.
Language:  English
Descriptors: Arabidopsis thaliana; Gene expression; Cloning;Saccharomyces cerevisiae; Potassium; Nucleotide sequences;Amino acid sequences
Abstract:  A membrane polypeptide involved in K+ transport ina higher plant was cloned by complementation of a yeast mutantdefective in K+ uptake with a complementary DNA library fromArabidopsis thaliana. A 2.65-kilobase complementary DNAconferred ability to grow on media with K+ concentration inthe micromolar range and to absorb K+ (or (86)Rb+) at ratessimilar to those in wild-type yeast. The predicted amino acidsequence (838 amino acids) has three domains: a channelforming region homologous to animal K+ channels, a cyclicnucleotide-binding site, and an ankyrin-like region.
17                                  NAL Call. No.: QH426.C8Cloning and expression of a chitinase gene from thehyperparasitic 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.  Includesreferences.
Language:  English
Descriptors: Deuteromycotina; Fungal antagonists; Fusariumoxysporum; 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 anAphanocladium album chitinase-overproducing mutant strain wereisolated by screening with antiserum against a 39 kDachitinase purified from this hyperparasitic fungus. Analysisof the isolated positive clones indicated that most of themcarried the same cDNA. A cDNA from this group was used as ahybridization probe to isolate an 8 kb DNA fragment from agenomic library of the wild-type strain. The chitinase 1 genewas mapped to this fragment by two independent approaches. Itspartial DNA sequence was in perfect agreement with an aminoterminal peptide sequence obtained by sequencing 23 aminoacids of the 39 kDa chitinase. Its transfer in Fusariumoxysporum resulted in a transformant producing both a proteinof about 39 kDa that cross-reacted with the chitinaseantiserum and a chitinase activity that was inhibited by thesame antiserum. Northern blot analysis indicates that thecloned chitinase gene was subject to catabolite repression andappeared inducible by chitin.
18                                  NAL Call. No.: 381 J824The cloning and expression of a gene encoding old yellowenzyme from Saccharomyces carlsbergensis.Saito, K.; Thiele, D.J.; Davio, M.; Lockridge, O.; Massey, V.Baltimore, Md. : American Society for Biochemistry andMolecular 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; Nucleotidesequences; Amino acid sequences
Abstract:  We have identified a gene that encodes Old YellowEnzyme in brewer's bottom yeast. The open reading frameencodes a polypeptide of 400 amino acids with Mr = 45,021.Using the T7 RNA polymerase system, recombinant enzyme wasexpressed in Escherichia coli. 17 mg of Old Yellow Enzyme wasobtained from a 3-liter cell culture, and the recombinantenzyme had NADPH oxidase activity. On fast protein liquidchromatography separation, the recombinant enzyme showed asingle large peak, while native enzyme from brewer's bottomyeast separated into five fractions on fast protein liquidchromatography. Southern blot analysis showed that there areat least two Old Yellow Enzyme genes in brewer's bottom yeastgenomic DNA. These results suggest that the heterogeneity ofOld Yellow Enzyme in Saccharomyces carlsbergensis is due tothe presence of multiple genes.
19                                  NAL Call. No.: 381 J824Cloning and expression of a yeast protein tyrosinephosphatase. Guan, K.; Deschenes, R.J.; Qiu, H.; Dixon, J.E.Baltimore, Md. : American Society for Biochemistry andMolecular Biology; 1991 Jul15.The Journal of biological chemistry v. 266 (20): p.12964-12970; 1991 Jul15. Includes references.
Language:  English
Descriptors: Saccharomyces cerevisiae; Tyrosine; Phosphoricmonoester hydrolases; Phosphorylation; Polymerase chainreaction; Cloning; Nucleotide sequences; Amino acid sequences
Abstract:  To study the regulation of tyrosinephosphorylation/dephosphorylation in Saccharomyces cerevisiae,a protein tyrosine phosphatase (PTPase) was cloned by thepolymerase chain reaction (PCR). Conserved amino acidsequences within the mammalian PTPases were used to designprimers which generated a yeast PCR fragment. The sequence ofthe PCR fragment encoded a protein with homology to themammalian PTPases. The PCR fragment was used to identify theyeast PTP1 gene which has an open reading frame encoding a335-amino acid residue protein. This yeast PTPase shows 26%sequence identity to the rat PTPase, although highly conservedresidues within the mammalian enzymes are invariant in theyeast protein. The yeast PTP1 is physically linked to the 5'-end of a heat shock gene SSB1. This yeast PTP1 gene wasexpressed in Escherichia coli and obtained in a highlypurified form by a single affinity chromatography step. Therecombinant yeast PTPase hydrolyzed phosphotyrosine containingsubstrates approximately 1000 times faster than aphosphoserine containing substrate. Gene disruption of yeastPTP1 has no visible effect on vegetative growth.
20                                  NAL Call. No.: QP501.E8Cloning and sequencing of arg3 and agr11 genes ofSchizosaccharomyces pombe on a 10-kb DNA fragment.Heterologous expression and mitochondrial targeting of theirtranslation 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; 1992Apr.  Includes references.
Language:  English
Descriptors: Yeasts; Saccharomyces cerevisiae; Mutants;Strains; Arginine; Anabolism; Genes; Genetic code; Geneexpression; Clones; Promoters; Nucleotide sequences; Aminoacid sequences; Acetylglutamic acid; Kinases; Oxidoreductases;Ornithine carbamoyltransferase; Enzyme activity; Translation;Mitochondria
Abstract:  The Schizosaccharomyces pombe arginine anabolicgenes encoding ornithine carbamoyltransferase (arg3) andacetylglutamate kinase/acetylglutamyl-phosphate reductase(arg11) were cloned by functional complementation of S. pombearg3 and arg11 mutant strains from S. pombe DNA genomiclibraries. Restriction analysis and sequencing of the twoclones showed that both genes are located on a common DNAfragment. The arg3 gene encodes a 327-amino-acid polypeptidepresenting a strong identity to Saccharomyces cerevisiae andhuman ornithine carbamoyltransferases. The arg11 gene encodesa 884-amino-acid polypeptide. The acetylglutamate kinase andacetylglutamate-phosphate reductase domains have been definedby their identity with the S. cerevisiae ARG5,6 protein. Thecloned arg11 gene from S. pombe does not complement an arg5,6mutation in S. cerevisiae, nor does the ARG5,6 gene complementthe S. pombe arg11- mutation. In contrast, both ornithinecarbamoyltransferase-encoding genes function in S. pombe.However, the S. pombe arg3 gene complements only weakly anarg3 S. cerevisiae strain, which is in agreement with the lowlevel of expression of the S. pombe gene in S. cerevisiae. Thesubcellular localization of both ornithinecarbamoyltransferases in the two yeasts indicates that, incontrast to the S. pombe enzyme, more than 95% of the S.cerevisiae enzyme remains in the S. pombe cytoplasm. The lowexpression 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 strikingsimilarities among themselves nor with the promoters of theequivalent genes of S. cerevisiae.
21                                NAL Call. No.: QH442.A1G4Cloning, expression and characterization of a cDNA encoding alipase 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 inEscherichia coli using poly(A)-selected mRNA from the fungus,Rhizopus (Rp.) delemar. Lipase-producing members of thelibrary were identified by means of a phenotypic score whereinthe release of fatty acids by lipase causes a characteristiccolor change in the growth medium. One such isolate containeda 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 substrateselectivity as the authentic fungal enzyme, hydrolyzing esterbonds at the stereospecific numbering (sn) sn-1 and sn-3, butnot the sn-2, positions of triglycerides. The completenucleotide sequence of the LIP cDNA was determined. Byreference to the N-terminal sequence of authentic Rp. delemarlipase, the lipase-encoding region was identified within thisfragment. The LIP cDNA encodes a putative preprolipaseconsisting of a 26-amino-acid(aa) signal sequence, a 97-aapropeptide, and a 269-aa mature enzyme. The predicted maturelipase has the same molecular weight and aa composition asthat of Rp. delemar, is highly homologous to that produced bythe fungus Rhizomucor miehei, and contains the consensuspentapeptide (Gly-Xaa-Ser-Yaa-Gly) which is conserved amonglipolytic enzymes. It is concluded that the LIP cDNA is anessentially full-length analogue of the lipase-encoding geneof Rp. delemar. The lipase encoded by the LIP cDNA occupies acytoplasmic location when synthesized in E. coli. Unprocessedforms of the lipase accumulate in E. coli.
22                                NAL Call. No.: QH442.A1G4Cloning, sequence analysis and transcriptional expression of aras 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; Nucleotidesequences; Transcription; Gene expression; Introns; Amino acidsequences; Developmental stages; Biological development; Camp; Mycelium
Abstract:  In the edible basidiomycete, Lentinus edodes, thepresence of a high level of intracellular cyclic AMP (cAMP) isclosely related to the onset of fruiting and/or primordiumformation. Since a close relationship between intracellularcAMP levels and expression of ras genes was reported fororganisms such as Saccharomyces cerevisiae and Dictyosteliumdiscoideum, we have cloned and sequenced a ras gene homologuefrom L. edodes (Le.), and analyzed its expression duringdevelopment of the fungus. This gene, named Le.ras, has acoding capacity of 217 amino acids (aa) interrupted by sixsmall introns. The deduced Le.Ras protein exhibited thehighest homology to the Schizosaccharomyces pombe RAS protein(219 aa): 86% homology in the N-terminal 80-aa sequence and74% homology in the next 80 aa. The Le.ras gene wastranscribed at similar levels during mycelial development infruiting-body formation, suggesting no direct correlation ofLe.ras expression with intracellular cAMP levels in thisorganism.
23                                  NAL Call. No.: QH426.C8Cloning, sequencing and expression of the Schwanniomycesoccidentalis 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.  Includesreferences.
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 Neurosporacrassa (am) have been shown to hybridize under reducedstringency conditions to genomic sequences of the yeastSchwanniomyces occidentalis. Using 5' and 3' gene-specificprobes, a unique 5.1 kb BclI restriction fragment thatencompasses the entire Schwanniomyces sequence has beenidentified. A recombinant clone bearing the unique BclIfragment has been isolated from a pool of enriched clones inthe yeast/E. coli shuttle vector pWH5 by colony hybridization.The identity of the plasmid clone was confirmed by functionalcomplementation of the Saccharomyces cerevisiae gdh-1mutation. The nucleotide sequence of the Schw. occidentalisGDH gene, which consists of 1380 nucleotides in a continuousreading frame of 459 amino acids, has been determined. Thepredicted amino acid sequence shows considerable homology withGDH proteins from other fungi and significant homology withall other available GDH sequences.
24                                  NAL Call. No.: 381 J824Cloning, structural analysis, and expression of the glycogenphosphorylase-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 andMolecular Biology; 1992 Feb05.The Journal of biological chemistry v. 267 (4): p. 2294-2302;1992 Feb05. Includes references.
Language:  English
Descriptors: Dictyostelium; Cell differentiation; Glycogenphosphorylase; Structural genes; Gene expression; Cloning;Enzyme activity; Purification; C-amp; Regulation; Nucleotidesequences; Amino acid sequences
Abstract:  The glycogen phosphorylase-2 (GP2) activity thatappears during the cell differentiation of Dictyostelium waspurified to homogeneity. The molecular weight of thenondenatured enzyme was 200,000 as determined by SephacrylS-300 gel filtration and was 107,000 on sodium dodecylsulfate-polyacrylamide gel electrophoresis, suggesting thatthe native enzyme consists of two similar subunits. The intactprotein was digested with trypsin and protease V8, and theresulting peptides were purified by microbore high pressureliquid chromatography. The peptides were sequenced, andoligonucleotides were constructed for polymerase chainreaction amplification of the GP2 gene from Dictyosteliumgenomic DNA template. The resulting polymerase chain reactionproducts were sequenced directly and were confirmed to encodeportions of the GP2 gene. These fragments were used to probe apartial EcoRI genomic library for the remainder of the GP2gene. The nucleotide sequence of the GP2-selected clonesrevealed an open reading frame of 2975 base pairs that wasinterrupted by two introns of 109 and 105 base pairs,respectively. The open reading frame encoded a protein of 992amino acids with a calculated molecular mass of 112,500 Da andan isoelectric point of 6.4. An unusual sequence within thesecond exon of GP2, in which the triplet CAA was repeated 11times, resulted in 11 in-frame glutamine residues of apossible 15 amino acids coded for by this region. The CAArepeat was transcribed, as shown by the sequence of cDNA.Comparison of the amino acid sequence of Dictyostelium GP2 tothe phosphorylases from other organisms revealed that theDictyostelium protein was 50 and 44% identical to yeast andrabbit muscle phosphorylases, respectively. Northern blotanalysis showed that GP2 mRNA was absent in amebas and theearly stages of development, reached a maximum level ofexpression at the slug stage, and then decreased in theterminal stages of development. Comparison of the mRNAexpression wi
25                                 NAL Call. No.: 442.8 G28The 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.  Includesreferences.
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 containrespectively the plus (P) and minus (M) mating-typeinformation in a transcriptionally silent state. Thatinformation is transposed from the mat2 or mat3 donor locusvia recombination into the expressed mating-type locus (mat1)resulting in switching of the cellular mating type. We haveidentified 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 oftranscription of the silent genes. Production of mRNA frommat3 is detectable in ch1 mutants. Furthermore, the ura4 geneinserted near mat3, weakly expressed in wild-type cells, isderepressed in clr1 mutants. The clr1 mutations also permitmeiotic recombination in the 15-kb mat2-mat3 interval, whererecombination is normally inhibited. The clr1 locus is in theright arm of chromosome II. We suggest that clr1 regulatessilencing of the mat2 and mat3 loci, and participates inestablishing the 'cold spot' for recombination by organizingthe chromatin structure of the mating-type region.
26                                 NAL Call. No.: 442.8 Z34Construction of a fusion gene comprising the Taka-amylase Apromoter and the Escherichia coli beta-glucuronidase gene andanalysis 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; Geneticengineering; Recombinant  DNA; Promoters; Alpha-amylase;Reporter genes; Beta-glucuronidase; Gene expression; Messenger RNA; Starch; Sugars; Carbohydrate metabolism; Genetictransformation
Abstract:  Northern blot analysis of glucose-grown and starchgrown mycelia of Aspergillus oryzae RIB40 was conducted usingthe cloned Taka-amylase A (TAA) gene as a probe. The amount ofmRNA homologous to the TAA gene was increased when this funguswas grown with starch as a sole carbon source. In order toanalyze the induction mechanism, we inserted the Escherichiacoli uidA gene encoding beta-glucuronidase (GUS) downstream ofthe TAA promoter and introduced the resultant fusion gene intothe A. oryzae genome. Production of a functional GUS proteinwas induced by starch, but not by glucose. When the effects ofvarious sugars on expression of the fusion gene were examined,the results suggested that the expression of the fusion genewas under control of the TAA gene promoter.
27                                 NAL Call. No.: QH450.C74Control 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; Literaturereviews
28                                   NAL Call. No.: 472 N21Coordination of expression of DNA synthesis genes in buddingyeast 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.  Includesreferences.
Language:  English
Descriptors: Saccharomyces; Gene expression; Genes; Dna;Synthesis
Abstract:  All of the DNA synthesis genes of budding yeastexamined so far are periodically expressed and hence undercell-cycle control. Expression occurs near the G1/S phaseboundary and the genes seem to be coordinately regulated. Theupstream promoter sequences of these genes have only a hexamerelement, ACGCGT (an MluI restriction site), in common. Here weshow that this hexamer is able to impart periodic expressionto a heterologous gene and, significantly, this expressionoccurs coincidentally with that of CDC9, one of the DNAsynthesis genes. We have also identified a protein that bindsspecifically to these sequences in a similar periodic manner.These ACGCGT sequences and the transcription factor that bindsto them therefore seem to be the elements controlling both theperiodic expression and coordinate regulation of the DNAsynthesis genes.
29                                  NAL Call. No.: 500 N21PCotranslational autoproteolysis involved in gene expressionfrom a double-stranded RNA genetic element associated withhypovirulence 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 UnitedStates of America v. 88 (4): p. 1167-1171; 1991 Feb15. Includes references.
Language:  English
Descriptors: Cryphonectria parasitica; Fungal diseases; Geneexpression; Genetic control; Hypovirulence; Nucleotidesequences; Proteolysis; Translation
30                                 NAL Call. No.: 442.8 Z34CYP1 (HAP1) is a determinant effector of alternativeexpression 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 isknown to activate a number of target genes in response to thepresence of heme. Several features of the protein, deducedfrom the sequence of the gene, suggest that CYP1 is a generalsensor of the redox state of the cell. To investigate furtherthe function of CYP1, we analysed its effects on thetranscription of two genes, HEM13 and 14DM, which arepreferentially expressed in anaerobiosis. HEM13 encodescoproporphyrinogen oxidase which catalyses the sixth enzymaticstep in the heme biosynthetic pathway and 14DM encodeslanosterol-14-demethylase which is involved in sterolbiosynthesis and is a member of the cytochrome P450 family.Isogenic CYP1+ and cyp1 deleted strains, either hemesufficient or heme-deficient (HEM1 disrupted), were grown inaerobic or anaerobic conditions, and transcripts of HEM13 and14DM were analysed on Northern blots. The results show that inanaerobic and in heme-deficient cells, CYP1 activates thetranscription of HEM13 and inhibits that of 14DM. Oppositeeffects of CYP1 are observed in aerobic heme-sufficient cells.We concluded that: (i) CYP1 is an efficient activatorespecially in heme-depleted cells; (ii) CYP1 exerts bothpositive and negative regulatory effects; (iii) the nature ofthe regulatory function of CYP1 depends on the target gene;and (iv) for a given gene, the presence or absence of heme oroxygen reverses the sense of CYP1-dependent regulation.
31                                 NAL Call. No.: 448.3 J82Developmental regulation of CUP gene expression through DNAmethylation in Mucor spp.Cano-Canchola, C.; Sosa, L.; Fonzi, W.; Sypherd, P.; RuizHerrera, J. Washington, D.C. : American Society forMicrobiology; 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 fromSaccharomyces cerevisiae or Mucor racemosus were used ashybridization probes to measure the methylation state andexpression of the CUP gene from Mucor rouxii at differentstages of growth. It was observed that the fungus contains aCUP multigene family. All the CUP genes were present in ahypermethylated DNA region in nongrowing and isodiametricallygrowing spores and were not transcribed at these stages. Aftergerm tube emergence, CUP genes became demethylated andtranscriptionally active. Development, demethylation, andtranscription of CUP genes were blocked by the ornithinedecarboxylase inhibitor 1,4-diaminobutanone. These resultssuggest that genes that are activated during developmentbecome demethylated in this fungus.
32                                NAL Call. No.: QH442.A1G4Dictyostelium discoideum as an expression host for thecircumsporozoite 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, Dictyosteliumdiscoideum (Dd), to express the Plasmodium falciparumcircumsporozoite protein (CS), a potential component of asubunit vaccine against malaria. This was accomplished via anexpression vector based on the discoidin I-encoding genepromoter, in which we linked a sequence coding for a Dd leaderpeptide to the almost complete CS coding region (pEDII-CS). CSproduction at both the mRNA and protein levels is induced bystarving cells in a simple phosphate buffer. Variation in pHor 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 CSaccumulation level. The CS produced in Dd seems similar to thenatural parasite protein as judged by its size and epitoperecognition by a panel of monoclonal antibodies. Weconstructed a second expression vector in which the CS isunder the control of a Dd ras promoter. CS accumulation canthen be induced by external addition of cAMP. Such a tightlyregulated promoter may allow expression of proteinspotentially toxic to the cell. Thus, Dd could be a usefuleukaryotic system to produce recombinant proteins, inparticular from human or animal parasites like P. falciparum.
33                                NAL Call. No.: 448.3 J823Differential gene expression and evidence of selectivetranslation during anaerobic germination of Mucor racemosussporangiospores. 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 Mucorracemosus develop into yeasts, whereas aerobically-germinatedsporangiospores become hyphae. Anaerobic germination was foundto have the following traits in common with the previouslycharacterized aerobic system of development: (i) immediate andvigorous protein synthesis upon exposure of the spores toliquid medium; (ii) a complete absence of RNA synthesis forthe first 20 min of germination; and (iii) a mobilization ofribosomal subunits into active polyribosomes. The proteinssynthesized in both systems during this 20 min interval mustbe specified by the pre-formed stable mRNA known to be storedin the dormant spore. The population of proteins manufacturedearly in anaerobic germination differed considerably from theset of proteins synthesized during the equivalent interval inair, suggesting that some unknown mechanism of selectivetranslation must operate. A few dozen of the more prominentproteins could be categorized according to their patterns ofsynthesis during germination. This should allow future work tofocus upon those genes and their products most closely linkedto development. The most promising candidates include severalproteins that are most conspicuous in the mature yeast and areamong those proteins selectively translated in large amountsfrom stored mRNA templates during anaerobic germination.
34                                NAL Call. No.: QD341.A2N8Direct introduction and transient expression of capped andnon-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 RNAmolecules into yeast spheroplasts. Plasmids containing thefirefly luciferase coding region were transcribed to yieldRNAs suitable for introduction into yeast cells and directassay of their translation products. The 5' noncoding regionsof the RNAs were derived either from the 5' noncoding regionsof firefly luciferase, poliovirus, or yeast-like-particle(VLP) L-A or M1 RNAs. Capped and noncapped MRNAs were made byT7 RNA polymerase-directed transcription and introduced intoyeast spheroplasts. The peak time of luciferase transientexpression from introduced RNAs was 2 - 4 h after theirintroduction. in contrast, transient expression of luciferasefrom a non-replicative, luciferase-encoding plasmid introducedinto the cells was maximal at 16 h. For capped mRNAs,luciferase activity increased linearly with transcript amountfor both yeast and human (HeLa) cells. Although non-cappedluciferase mRNAs were expressed more efficiently followingintroduction into yeast than into HeLa cells, the 5' noncodingsequences from yeast double-stranded (dS)RNA VLP RNAsconferred no greater apparent cap-independence than non-VLPRNA sequences in this transient expression assay. The RNAtransient expression system will allow the study oftranslation of capped and non-capped RNAs in yeast cells andof the replicative cycle of yeast virus-like RNA genomes.
35                                NAL Call. No.: 442.8 J828Domain structure in actin-binding proteins: expression andfunctional 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; Plantproteins; Mutants; Gene expression; Protein synthesis; Bindingsite; 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 ofactin filaments. Sequence comparison with functionally relatedproteins, such as gelsolin, villin, or fragmin revealed highlyconserved domains which are thought to be of functionalsignificance. To attribute the different activities of theseverin molecule to defined regions, progressively truncatedseverin polypeptides were constructed. The complete cDNAcoding for 362 (DS362) amino acids and five 3' deletionscoding 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 thencharacterized with respect to their effects on thepolymerization or depolymerization kinetics of G- or F-actinsolutions and their binding to G-actin. Furthermore, the Ca2+binding of these proteins was investigated with a 45Ca-overlayassay and by monitoring Ca2+-dependent changes in tryptophanfluorescence. Bacterially expressed DS362 showed the sameCa2+-dependent activities as native severin. DS277, missingthe 85 COOH-terminal amino acids of severin, had lost itsstrict Ca2+ regulation and displayed a Ca2+-independentcapping activity, but was still Ca2+ dependent in its severingand nucleating activities. DS151 which corresponded to thefirst domain of gelsolin or villin had completely lostsevering and nucleating properties. However, a residualsevering activity of approximately 2% was detectable if 26amino 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 seconddomain. Measuring the fluorescence enhancement of pyrenelabeled G-actin in the presence of DS111 showed that the firstactin-binding site was present in the NH2-terminal 111 aminoacids. Extension by six or more amino acids stabilized t
36                                  NAL Call. No.: 385 J822 Adominant mutation that alters the regulation of INO1expression 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; Geneexpression; Genetic regulation; Choline; Myo-inositol;Biosynthesis
Abstract:  A dominant, single nuclear gene mutation, CSE1,caused inositol auxotrophy in yeast cells. The inositolrequirement was marked when choline was present in the medium.Inositol-1-phosphate synthase, the regulatory enzyme ofinositol synthesis, is repressed by inositol, or moreprofoundly by a combination of inositol and choline in thewild type. In CSE1, the level of inositol-1-phosphate synthasewas low and was greatly repressed on the addition of cholinealone. In accordance with this, INO1 mRNA encoding the enzymewas low even under the derepressed conditions and wasprofoundly decreased by choline in CSE1. But in the wild type,the addition of choline alone had little effect. An INO1-lacZfusion was constructed and the control of the INO1 promoter inCSE1 was studied. lacZ expression was repressed not only byinositol, but also by choline in CSE1, whereas it wasrepressed by inositol, but only slightly by choline in thewild type. CSE1 was unlinked to the INO1 structural gene. ThusCSE1 was thought to be a regulatory mutation. Furthermore,when the CDP-choline pathway was mutationally blocked, cholinedid not affect IN01 expression, indicating that the metabolismof choline via the CDP-choline pathway is required for INO1repression.
37                                  NAL Call. No.: 23 AU792Effect of Kabatiella caulivora isolates and host growth stageon symptom expression and resistance in Trifoliumsubterraneum.Barbetti, M.J.; Gillespie, D.J.; Collins, W.J.East Melbourne : Commonwealth Scientific and IndustrialResearch 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.E8Efficient expression of bovine alpha-lactalbumin inSaccharomyces 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; 1991Dec.  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 inSaccharomyces cerevisiae under the control of the alpha-matingpromotor. Growth conditions were found to be critical for theexpression: recombinant alpha-lactalbumin could only bedetected in the medium provided the culture was grown atneutral pH. The secreted bovine alpha-lactalbumin isenzymatically active and identical to the whey protein, asconfirmed by SDS/PAGE, IEF, ultraviolet and CD spectralanalysis, and amino-terminal sequence determination.
39                                  NAL Call. No.: QH324.C7Enhanced survival of yeast expressing an antifreeze geneanalogue after freezing.McKown, R.L.; Warren, G.J.Orlando, Fla. : Academic Press; 1991 Oct.Cryobiology v. 28 (5): p. 474-482; 1991 Oct.  Includesreferences.
Language:  English
Descriptors: Saccharomyces cerevisiae; Bacterial proteins;Cold resistance; Cryoprotectants; Freezing; Gene expression;Genetic transformation; Nucleotide sequences; Survival
40                                  NAL Call. No.: QH442.B5Evaluation 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.  Includesreferences.
Language:  English
Descriptors: Saccharomyces cerevisiae; Genetic engineering;Genetic transformation; Gene expression; Immunoglobulinstructural genes; Mice; Genetic code; Protein synthesis
41                                NAL Call. No.: 442.8 J828Evolutionary conservation of the human nucleolar proteinfibrillarin 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; Cellultrastructure
Abstract:  NOP1 is an essential nucleolar protein in yeastthat is associated with small nucleolar RNA and required forribosome biogenesis. We have cloned the human nucleolarprotein, fibrillarin, from a HeLa cDNA library. Humanfibrillarin is 70% identical to yeast NOP1 and is also thefunctional homologue since either human or Xenopus fibrillarincan complement a yeast nop1(-) mutant. Human fibrillarin islocalized in the yeast nucleolus and associates with yeastsmall nucleolar RNAs. This shows that the signals withineucaryotic fibrillarin required for nucleolar association andnucleolar function are conserved from yeast to man. However,human fibrillarin only partially complements in yeastresulting in a temperature-sensitive growth, concomitantlyaltered rRNA processing and aberrant nuclear morphology. Asuppressor of the human fibrillarin ts-mutant was isolated andfound to map intragenically at a single amino acid position ofthe human nucleolar protein. The growth rate of yeast nop1(-)strains expressing Xenopus or human fibrillarin or the humanfibrillarin suppressor correlates closely with their abilityto efficiently and correctly process pre-rRNA. These findingsdemonstrate for the first time that vertebrate fibrillarinfunctions in ribosomal RNA processing in vivo.
42                                  NAL Call. No.: 381 J824Expression and characterization of a recombinant yeastisoleucyl-tRNA synthetase.Racher, K.I.; Kalmar, G.B.; Borgford, T.J.Baltimore, Md. : American Society for Biochemistry andMolecular 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 arecombinant Saccharomyces cerevisiae isoleucyl-tRNA synthetase(IRS) gene in Escherchia coli, as well as the purification andcharacterization of the recombinant gene product. High levelexpression of the yeast isoleucyl-tRNA synthetase gene wasfacilitated by site-specific mutagenesis. The putativeribosome-binding site of the yeast IRS gene was made to be theconsensus of many highly expressed genes of E. coli.Mutagenesis simultaneously created a unique BclI restrictionsite such that the gene coding region could be convenientlysubcloned as a "cassette." The variant gene was cloned intothe expression vector pKK223-3 (Brosius, J., and Holy, A.(1984) Proc. Natl. Acad. Sci. U.S.A. 81, 6929-6933) therebycreating the plasmid pKR4 in which yeast IRS expression isunder the control of the isopropyl-thio-beta-galactopyranoside(IPTG)-inducible tac promoter. Recombinant yeast IRS, on theorder of 10 mg/liter of cell culture, was purified from pKR4-infected and IPTG-induced E. coli strain TG2. Yeast IRS waspurified to homogeneity by a combination of anion-exchange andhydroxyapatite gel chromatography. Inhibition of yeast IRSactivity by the antibiotic pseudomonic acid A was tested. Theyeast IRS enzyme was found to be 10(4) times less sensitive toinhibition by pseudomonic acid A (Ki = 1.5 X 10(-5) M) thanthe E. coli enzyme. E. coli strain TG2 infected with pKR4, andinduced with IPTG, had a plating efficiency of 100% atinhibitor concentrations in excess of 25 micromole/ml. At thesame concentration of pseudomonic acid A, E. coli strain TG2infected with pKK223-3 had a plating efficiency < 1%. Theability of yeast IRS to rescue E. coli from pseudomonic acid Asuggests that the eukaryotic synthetase has full activity inits prokaryotic host and has specificity for E. coli tRNAile.
43                                   NAL Call. No.: 381 AR2Expression and function of heterologous forms of malatedehydrogenase 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.I628Expression and secretion in yeast of active insect defensin,an inducible antibacterial peptide from the fleshfly Phormiaterranovae. 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; Antibacterialproperties; Gene expression; Saccharomyces cerevisiae;Nucleotide sequences; Amino acid sequences
45                                NAL Call. No.: 448.3 J823Expression 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; Proteinsecretion; 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 induciblepromoter. OmpA-nuclease and lipo-nuclease contain the maturestaphylococcal nuclease sequence preceded by the Escherichiacoli OmpA and lipoprotein signal sequences, respectively,whereas Pin-nuclease lacks a defined signal sequence at itsamino terminus. We found that: (a) the nuclease gene productssynthesized in yeast are active, but they do not affect cellgrowth; b) OmpA-nuclease and lipo-nuclease are partiallyprocessed and constitute approximately 1.0-1.5% of the yeastcell protein; (c) OmpA and lipoprotein signal sequencesfunction similarly in secretion, allowing 35-40% of theprocessed nuclease to be translocated into the yeastperiplasm; and (d) Pin-nuclease, which lacks hydrophobicsequences at its amino-terminus, is accumulated at a leveltenfold lower than the hybrid proteins that do contain signalsequences. Nevertheless, 50% of the enzyme activity of Pinnuclease in yeast is localized in the periplasmic space.
46                                  NAL Call. No.: QH426.C8Expression and sequence comparison of the Aspergillus nigerand Aspergillus tubigensis genes encoding polygalacturonaseII.Bussink, H.J.D.; Buxton, F.P.; Visser, J.Berlin, W. Ger. : Springer International; 1991.Current genetics v. 19 (6): p. 467-474; 1991.  Includesreferences.
Language:  English
Descriptors: Aspergillus niger; Aspergillus; Gene expression;Genetic code; Polygalacturonase; Nucleotide sequences;Restriction mapping; Amino acid sequences
Abstract:  The structure and expression of thepolygalacturonase-encoding pgaII genes of two recentlyrecognized species, Aspergillus niger and Aspergillustubigensis, was investigated. While the structure of the pgaIIgenes is very similar, showing 83% DNA sequence identity and94% identity at the amino acid level, they have divergedsignificantly. The NH2-terminal sequence suggests that thesePGs are made as pre pro-proteins and the secretory propeptideof the PGII precursors shows sequence homology with some otherfungal pro-peptides. The expression of the pgaII genes isstrongly regulated by the carbon source and the A. tubigensisgene is expressed and regulated in A. niger transformants. Thelow similarity of the fungal PGs with those of bacterial andplant origin is discussed in relation to the possiblefunctional role of specific amino acids.
47                                 NAL Call. No.: 448.3 J82Expression in Escherichia coli of the Saccharomyces cerevisiaeCCT gene encoding cholinephosphate cytidylyltransferase.Tsukagoshi, Y.; Nikawa, J.; Hosaka, K.; Yamashita, S.Washington, D.C. : American Society for Microbiology; 1991Mar. Journal of bacteriology v. 173 (6): p. 2134-2136. ill;1991 Mar.  Includes references.
Language:  English
Descriptors: Saccharomyces cerevisiae; Transferases; Geneticcode; Cloning; Plasmids; Escherichia coli; Enzyme activity;Gene expression
Abstract:  The coding region of the CCT gene from the yeastSaccharomyces cerevisiae was cloned into the pUC18 expressionvector. The plasmid directed the synthesis of an activecholinephosphate cytidylyltransferase in Escherichia coli,confirming that CCT is the structural gene for this enzyme.The enzyme produced in E. coli efficiently utilizedcholinephosphate and N,N-dimethylethanolaminephosphate, but Nmethylethanolaminephosphate and ethanolaminephosphate werepoor substrates. Consistently, disruption of the CCT locus inthe wild-type yeast cells resulted in a drastic decrease inactivities with respect to the former two substrates. Whenactivity was expressed in E. coli, over 90% was recovered inthe cytosol, whereas most of the activity of yeast cells wasassociated with membranes, suggesting that yeast cells possessa mechanism that promotes membrane association ofcytidylyltransferase.
48                                 NAL Call. No.: 442.8 Z34Expression in yeast of a cDNA copy of the K2 killer toxingene. Dignard, D.; Whiteway, M.; Germain, D.; Tessier, D.;Thomas, D.Y. Berlin, W. Ger. : Springer International; 1991May.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; Inducedmutations; Mutants; Enzyme activity; Proteinases; Geneexpression
Abstract:  A cDNA copy of the M2 dsRNA encoding the K2 killertoxin of Saccharomyces cerevisiae was expressed in yeast usingthe yeast ADH1 promoter. This construct produced K2-specifickilling and immunity functions. Efficient K2-specific killingwas dependent on the action of the KEX2 endopeptidase and theKEX1 carboxypeptidase, while K2-specific immunity wasindependent of these proteases. Comparison of the K2 toxinsequence with that of the K1 toxin sequence shows thatalthough they share a common processing pathway and are bothencoded by cytoplasmic dsRNAs of similar basic structure, thetwo toxins are very different at the primary sequence level.Site-specific mutagenesis of the cDNA gene establishes thatone of the two potential KEX2 cleavage sites is critical fortoxin action but not for immunity. Immunity was reduced by aninsertion of two amino acids in the hydrophobic amino-terminalregion which left toxin activity intact, indicating anindependence of toxin action and immunity.
49                                 NAL Call. No.: QP501.B64Expression of a cDNA clone encoding the haem-binding domain ofChlorella 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; Nucleotidesequences; Spectral analysis
Abstract:  A partial cDNA clone coding for the haem-bindingdomain of NADH:nitrate reductase (EC 1.6.6.1) (NR) from theunicellular green alga Chlorella vulgaris has been isolated,sequenced and expressed. A 1.2 kb cDNA (pCVNR1) was isolatedfrom a lambda gt 11 expression library produced frompolyadenylated RNA extracted from nitrate-grown Chlorellacells. pCVNR1 hybridized to a 3.5 kb mRNA transcript that wasnitrate-inducible and absent from ammonium-grown cells. Theentire sequence of pCVNR1 was obtained and found to have asingle uninterrupted reading frame. The derived amino acidsequence of 318 amino acids has a 45-50 % similarity tohigher-plant NRs, including Arabidopsis thaliana, spinach(Spinacia oleracea) and tobacco (Nicotiana tabacum). Acomparison with the putative domain structure of higher-plantnitrate reductases suggested that this sequence contains thecomplete haem-binding domain, approximately one-third of theMo-pterin domain and no FAD-binding domain. A 32% sequencesimilarity is evident when comparing the Chlorella NR haemdomain with that of calf cytochrome b5. Expression of pCVNR1in a pET vector synthesized a 35 kDa protein that wasantigenic to anti-(Chlorella NR) antibody. The spectralproperties of this protein (reduced and oxidized) in the400-600 nm region are identical with those of native ChlorellaNR and indicate that haem is associated with the protein.
50                                 NAL Call. No.: 442.8 Z34Expression of a dominant negative allele of cdc2 preventsactivation 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; Dominantlethals; Protein kinase; Serine; Enzyme activity; Celldivision; Phosphoproteins; Amino acid sequences; Nucleotidesequences; Induced mutations
Abstract:  The cdc2 gene of the fission yeastSchizosaccharomyces pombe encodes a 34 kDa phosphoprotein withserine/threonine protein kinase activity that acts as the keycomponent in regulation of the eukaryotic cell cycle. We useda repressible promoter fused to the cdc2 cDNA to isolateconditionally dominant negative mutants of cdc2. One of thesemutants, DL5, is described in this paper. Overexpression ofthe mutant protein in a wild-type cdc2 background is lethaland confers cell cycle arrest with a typical cdc-phenotype.Sequencing of the mutant cdc2 gene revealed a single aminoacid substitution in a region highly conserved in cdc2-likeproteins. The mutant protein exhibits no protein kinaseactivity, but is able to bind a component(s) required for anactive protein kinase complex and thereby prevents binding ofthis component(s) to the co-existing wild-type cdc2 protein.We also demonstrate that S. pombe p34(cdc2) contains nophosphoserine.
51                                  NAL Call. No.: 450 P692Expression of a fungal sesquiterpene cyclase gene intransgenic tobacco. Hohn, T.M.; Ohlrogge, J.B.Rockville, Md. : American Society of Plant Physiologists; 1991Sep. Plant physiology v. 97 (1): p. 460-462; 1991 Sep. Includes references.
Language:  English
Descriptors: Nicotiana tabacum; Transgenics; Fusariumsporotrichioides; Gene expression; Biosynthesis;Sesquiterpenes; Ligases; Enzyme activity; Disease resistance
Abstract:  The complete coding sequence for the trichodienesynthase gene from Fusarium sporotrichioides was introducedinto tobacco (Nicotiana tabacum) under the regulation of thecauliflower mosaic virus 35S promoter. Expression oftrichodiene synthase was demonstrated in the leaves oftransformed plants. Leaf homogenates incubated with[3H]farnesyl pyrophosphate produced trichodiene as a majorproduct. Trichodiene was detected in the leaves of atransformed plant at a level of 5 to 10 nanograms per gramfresh weight. The introduction of a fungal sesquiterpenecyclase gene into tobacco has resulted in the expression of anactive enzyme and the accumulation of low levels of itssesquiterpenoid product.
52                                  NAL Call. No.: TX393.J6Expression of a wheat gliadin protein in yeast (Saccharomycescerevisiae). 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; Saccharomycescerevisiae
Abstract:  A full-length CDNA encoding a gamma-type gliadinfrom wheat was expressed in Saccharomyces cerevisiae. A yieldof 1-2% of total cell protein was obtained, and the level ofexpression was not affected by using a protease-deficientstrain of yeast. Analysis of the purified gamma-gliadin showedthat cleavage of the signal peptide had occurred but thatincorrect disulphide bond formation had resulted in a range ofoligomers and polymers in addition to monomers.
53                                 NAL Call. No.: 448.3 J82Expression of bacterial mercuric ion reductase inSaccharomyces cerevisiae. Rensing, C.; Kues, U.; Stahl, U.;Nies, D.H.; Friedrich, B. Washington, D.C. : American Societyfor 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 ionreductase was cloned under the control of the yeast promoterfor alcohol dehydrogenase I in the yeast-Escherichia colishuttle plasmid pADH040-2 and transformed into Saccharomycescerevisiae AH22. The resulting transformant harbored stablecopies of the merA-containing hybrid plasmid, displayed afivefold increase in the MIC of mercuric chloride, andsynthesized mercuric ion reductase activity.
54                                 NAL Call. No.: SB599.P45Expression of disease resistance response genes in nearisogenic pea cultivars following challenge by Fusariumoxysporum 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; Geneexpression; Dominance; Genes; Varietal susceptibility; Races;Host parasite relationships
Abstract:  Previous research on the pea/Fusarium oxysporumsystem has shown that the internal invasion of tap roots andhypocotyls was less frequent in the cultivar Vantage, whichpossesses a single dominant gene for resistance to Race 1,than in the cultivar M410 which is susceptible. We monitoredthese cultivar tissues for the expression of diseaseresistance response genes 49 and 206, previously associatedwith the expression of non-host resistance, in peas grown insoil naturally infested with Fusarium oxysporum f. sp. pisi(Race 1). The accumulation of distinct RNA products homologouswith both genes was evident in the resistant cultivar in aperiod prior to and during the onset of wilt symptoms in thesusceptible cultivar. Additional periods of RNA accumulationoccurred in older flowering seedlings of both cultivars. Thesewere not gene-specific RNA accumulations and were probablyunrelated to resistance. Thus these response genes maycontribute to normal cellular functions which are alsoutilized in the disease resistance response. The biochemicalfunctions of genes 49 and 206 remain unknown.
55                                 NAL Call. No.: QK600.B72Expression 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 moleculargenetics of fungi / edited by J. F. Peberdy, C. E. Caten, J.E. Ogden and J. W. Bennett. Symposium of the BritishMycological 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; Genetictransformation; Proteins; Structural genes; Vectors; Plasmids;Literature reviews
56                                NAL Call. No.: QH442.A1G4Expression of lacZ gene fusions affects downstreamtranscription 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 lacZsequences are often used to characterize gene expression inyeast cells. By Northern analysis, we found that such genesproduce multiple transcripts due to inefficient 3'-endformation. The same transcript pattern was found for tworelated chimeric genes when these genes were cloned separatelyinto the commonly used vector, YIp5, and integrated into theyeast genome at two different locations. Each chimeric genewas composed of promoter and N-terminal coding regions fromthe yeast SSA1 or SSA2 genes fused in-frame to the lac operon.Transcripts were shown to initiate within the yeast promoterfragment, but transcript size indicated that 3' ends werelocalized to three different regions: within the lac operonnear the 3' end of the lacZ gene; near a terminator regionpreviously identified upstream of the URA3 gene in YIp5; andat the URA3 terminator region. Readthrough transcription ofthe URA3 promoter from upstream lac sequences decreased thebasal activity of the URA3 promoter, although induced URA3transcription levels were unaffected. This readthroughtranscription also resulted in a novel, longer URA3transcript.
57                                NAL Call. No.: QR360.A1J6Expression of potyvirus coat protein in Escherichia coli andyeast 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 thepotyvirus, Johnsongrass mosaic virus (JGMV), was expressed inEscherichia coli or yeast, it assembled to form potyvirus-likeparticles. The particles were heterogeneous in length with astacked-ring appearance and resembled JGMV particles in theirflexuous morphology and width. This cell-free assembly systemshould permit analysis of the mechanisms of particle assemblyand genome encapsidation. Two mutant forms of CP produced bysite-directed mutagenesis failed to assemble into virus-likeparticles.
58                                  NAL Call. No.: 500 N21PExpression of RNase P RNA in Saccharomyces cerevisiae iscontrolled 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 UnitedStates of America v. 88 (16): p. 6986-6990; 1991 Aug15. Includes references.
Language:  English
Descriptors: Saccharomyces cerevisiae; Strains; Geneexpression; Genetic code; Nucleotide sequences; Promoters;Ribonucleases; Rna polymerase; Transcription
Abstract:  The RNA subunit of Saccharomyces cerevisiae nuclearRNase P is encoded by a single-copy, essential gene, RPR1. The369-nucleotide mature form of the RNA has an apparentprecursor with an 84-nucleotide 5' leader and approximately 33nucleotides of additional 3' sequence. Analysis of RPR1transcription in a strain with a temperature-sensitive lesionin RNA polymerase III shows that the gene is transcribed invivo by RNA polymerase III. Examination of potential promoterregions using both progressive upstream deletions and pointmutations indicates that at least two sequences containedwithin the 5' leader region are essential for expression invivo, while sequences farther upstream influence efficiency.The required leader elements resemble tRNA gene-like A-box andB-box internal promoters in sequence and spacing. As in thetRNA genes, transcription factor TFIIIC binds to this regionin vitro and binding is severely reduced by either A-box or Bbox point mutations that impair expression in vivo. It thusappears that the yeast RNase P RNA gene has adopted a promoterstrategy that places an RNA polymerase III "internal" promoterupstream of the mature structural domain to help drivetranscription.
59                                  NAL Call. No.: 385 J822Expression of RNase Rh from Rhizopus niveus in yeast andcharacterization 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; Nucleotidesequences; Amino acid sequences
60                                 NAL Call. No.: 448.3 AP5Expression of the Escherichia coli beta-glucuronidase gene inPseudocercosporella herpotrichoides.Bunkers, G.J.Washington, D.C. : American Society for Microbiology; 1991Oct. Applied and environmental microbiology v. 57 (10): p.2896-2900; 1991 Oct. Includes references.
Language:  English
Descriptors: Pseudocercosporella herpotrichoides; Geneticanalysis; Genetic transformation; Escherichia coli; Betaglucuronidase; Enzyme activity; Gene expression; Geneticmarkers; Plant pathogens; Hordeum vulgare; Triticum aestivum;Secale cereale
Abstract:  The plant-pathogenic fungus Pseudocercosporellaherpotrichoides has been successfully transformed by using twodifferent positive selection systems in combination with theEscherichia coli gusA gene. The selectable markers used inthis study were the hygromycin B phosphotransferase gene (hph)from E. coli and the gene (bml) for beta-tubulin from abenomyl-resistant mutant of Neurospora crassa. A lowertransformation rate was obtained with the bml system than withthe hph system. Conversely, cotransformation frequencies, asdetermined with medium plates containing the chromogenicsubstrate 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 mitoticallystable, and both the selectable gene and gusA were maintainedthrough conidiation. The vector DNA was integrated into thegenome, and the number and sites of insertion varied amongtransformants. Enzyme assays of mycelial extracts showed thatbeta-glucuronidase activity was highest in transformants witha high gusA copy number. Expression of gusA during growth ofthe fungus on plants was easily detectable and did not affectpathogenicity. These results form the basis for constructionof a versatile and sensitive reporter gene system for P.herpotrichoides.
61                                  NAL Call. No.: 381 J824Expression of the yeast plasma membrane [H+]ATPase insecretory vesicles. A new strategy for directed mutagenesis.Nakamoto, R.K.; Rao, R.; Slayman, C.W.Baltimore, Md. : American Society for Biochemistry andMolecular 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; Secretorygranules; Enzyme activity; Transport processes
Abstract:  Secretory vesicles that accumulate in thetemperature-sensitive sec6-4 strain of yeast have been shownto contain a vanadate-sensitive ATPase, presumably en route tothe plasma membrane (Walworth, N. C., and Novick, P. J. (1987)J. Cell Biol. 105, 163-174). We have now established thisenzyme to be a fully functional form of the PMA1 [H+]ATPase,identical in its catalytic properties to that found in theplasma membrane. In addition, the secretory vesicles aresealed tightly enough to permit the measurement of ATPdependent proton pumping with fluorescent probes. We have goneon to exploit the vesicles as an expression system for sitedirected mutants of the ATPase. For this purpose, a sec6-4strain has been constructed in which the chromosomal PMA1 geneis under control of the GAL1 promoter; the mutant pma1 alleleto be studied is introduced on a centromeric plasmid under thecontrol of a novel heat shock promoter. In galactose medium at23 degrees C, the wild-type ATPase is produced and supportsnormal vegetative growth. When the cells are switched toglucose medium at 37 degrees C, however, the wild-type geneturns off, the mutant gene turns on, and secretory vesiclesaccumulate. The vesicles contain a substantial amount of newlysynthesized, plasmid-encoded ATPase (5-10% of total vesicleprotein), but only traces of residual wild-type PMA1 ATPaseand no detectable mitochondrial ATPase, vacuolar ATPase, oracid or alkaline phosphatase. To test the expression strategy,we have made use of pma1-105 (Ser 368 leads to Phe), avanadate-resistant mutant previously characterized by standardmethods (Perlin, D. S., Harris, S. L., Seto-Young, D., andHaber, J. E. (1989) J. Biol. Chem. 264, 21857-21864). Insecretory vesicles, as expected, the plasmid-borne pma1 allele105 gives rise to a mutant enzyme with a reduced rate of ATPhydrolysis and a 100-fold increase in Ki for vanadate. Protonpumping is similarly resistant to vanadate. Thus, the vesiclesappear well suited f
62                                  NAL Call. No.: 500 N21PExpression of three mammalian cDNAs that interfere with RASfunction 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 UnitedStates of America v. 88 (7): p. 2913-2917. ill; 1991 Apr01. Includes references.
Language:  English
Descriptors: Saccharomyces cerevisiae; Strains; Yeasts; Aminoacid sequences; Cloning; Dna; Gene expression; Genetic code;Heat shock; Nucleotide sequences; Oncogenes
Abstract:  Saccharomyces cerevisiae strains expressing theactivated RAS2(Val19) gene or lacking both cAMPphosphodiesterase genes, PDE1 and PDE2, have impaired growthcontrol and display an acute sensitivity to heat shock. Wehave isolated two classes of mammalian cDNAs from yeastexpression libraries that suppress the heat shock-sensitivephenotype of a RAS2(Val19) strain. Members of the first classof cDNAs also suppress the heat shock-sensitive phenotype ofpde1- pde2- strains and encode cAMP phosphodiesterases.Members of the second class fail to suppress the phenotype ofpde1- pde2- strains and therefore are candidate cDNAs encodingproteins that interact with RAS proteins. We report thenucleotide sequence of three members of this class. Two ofthese cDNAs share considerable sequence similarity, but noneare clearly similar to previously isolated genes.
63                                 NAL Call. No.: 442.8 Z34Expression of variant nuclear Arabidopsis tRNA(Ser) genes andpre-tRNA maturation differ in HeLa, yeast and wheat germextracts. 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; Geneexpression; 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 inthe Arabidopsis nuclear genome. One tRNA(Ser) (AGA) gene andtwo tRNA(Tyr) (GTA) genes occur in tandem arrangement on a 1.5kb unit that is amplified about 20-fold at a singlechromosomal site. Here we have studied the in vitro expressionof 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 havein part adverse effects on the expression of these genes indifferent cell-free systems: (i) C10 and A62 in planttRNA(Ser) genes, which correspond to G10 and C62,respectively, in all known vertebrate tRNA genes, result in areduced transcription efficiency in HeLa but not in yeastextract. This indicates that yeast RNA polymerase IIItolerates nucleotide substitutions at positions 10 [5'internal control region (ICR)] and 62 (3' ICR), whereas thevertebrate RNA polymerase III requires a more stringentconsensus sequence. (ii) Processing of a pre-tRNA(Ser) with amismatch in the aminoacyl stem is impaired in HeLa, yeast andwheat germ extracts, however, a mismatch in the anticodon stemis deleterious for HeLa and wheat germ but not for yeastprocessing enzymes. The unexpectedly high number of potentialtRNA(Ser) pseudogenes in the cluster--quite in contrast to thetRNA(Tyr) genes which mainly code for functional tRNAs--suggested that tRNA(Ser) (AGA) genes also occur elsewhere inthe genome. We present evidence that single copies oftRNA(Ser) (AGA) genes do indeed exist outside the tRNA genecluster.
64                                 NAL Call. No.: 442.8 Z34Expression of yeast cytochrome c1 is controlled at thetranscriptional 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 Saccharomycescerevisiae (CYT1) was localized on chromosome XV. Its upstreamregion was identified by functional complementation. Fusion tothe lacZ reporter gene on a CEN plasmid allowed study of theeffect of carbon sources and of specific deletion mutations onexpression of the gene in yeast transformants. Detailedpromoter analysis combined with expression studies inrecipient strains defective in regulatory genes identifiedcis-acting sites and transcription factors involved in theregulated expression of the cytochrome c1 gene. These analysesshowed that, in the presence of glucose, transcription of CYT1is positively controlled by oxygen, presumably through thehaem signal, and mediated by the HAP1-encoded transactivator.It is additionally regulated by the HAP2/3/4 complex whichmediates gene activation mainly under glucose-free conditions.Basal transcription is, in part, effected by CPF1, acentromere and promoter-binding factor.
65                                  NAL Call. No.: QR360.J6Expression of yeast L-A double-stranded RNA virus proteinsproduces derepressed replication: a ski- phenocopy.Wickner, R.B.; Icho, T.; Fujimura, T.; Widner, W.R.Washington, D.C. : American Society for Microbiology; 1991Jan. 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 RNAvirus of Saccharomyces cerevisiae has two large open readingframes, ORF1, which encodes the major coat protein, and ORF2,which encodes a single-stranded RNA-binding protein having asequence diagnostic of viral RNA-dependent RNA polymerases.ORF2 is expressed only as a Gag-Pol-type fusion protein withORF1. We have constructed a plasmid which expresses theseproteins from the yeast PGK1 promoter. We show that thisplasmid 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 ORF2expression, providing a potential in vivo assay for the RNApolymerase and single-stranded RNA-binding activities of thefusion protein determined by ORF2. ORF1 expression, like ahost ski- mutation, can suppress the usual requirement of M1for the MAK11, MAK18, and MAK27 genes and allow a defective LA (L-A-E) to support M1 replication. These results suggestthat expression of ORF1 from the vector makes the cell a skiphenocopy. Indeed, expression of ORF1 in a wild-type killermakes it a superkiller, suggesting that a target of the SKIantiviral system may be the major coat protein.
66                                NAL Call. No.: QH442.A1G4 Anexpression vector for the phytopathogenic fungus, Ustilagomaydis. 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; Genetictransformation; Marker genes; Drug resistance; Hygromycin b;Dna; Cloning; Nucleotide sequences; Mycotoxins; Toxicity;Virus-like particles
Abstract:  We have constructed an expression vector for thephytopathogenic fungus Ustilago maydis. This vector, pUXV,expresses genes located downstream from a U. maydisglyceraldehyde-3-phosphate dehydrogenase promoter. PlasmidpUXV also contains a selective marker gene conferringresistance to the antibiotic hygromycin B and a U. maydisautonomously replicating sequence, UARS, allowing hightransformation efficiency. Expression of a cDNA from thetoxin-encoding region of the U. maydis virus P6 in pUXVresulted in as much killing activity as from viral particleswhen evaluated by killer plate assay. Plasmid pUXV preservesessential sequences from pUC12 and is therefore a shuttlevector for U. maydis and Escherichia coli.
67                                  NAL Call. No.: 500 N21PFunctional expression of a probable Arabidopsis thalianapotassium 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 UnitedStates 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 acidsequences; Cloning; Nucleotide sequences
Abstract:  We report the isolation of a cDNA (KAT1) fromArabidopsis thaliana that encodes a probable K+ channel. KAT1was cloned by its ability to suppress a K+ transport-defectivephenotype in mutant Saccharomyces cerevisiae cells. Thissuppression is sensitive to known K+ channel blockers,including tetraethylammonium and Ba2+ ions. The KAT1 cDNAcontains an open reading frame capable of encoding a 78-kDaprotein that shares structural features found in the Shakersuperfamily of K+ channels. These include a cluster of sixputative membrane-spanning helices (S1-S6) at the aminoterminus of the protein, a presumed voltage-sensing regioncontaining Arg/Lys-Xaa-Xaa-Arg/Lys repeats within S4, and thehighly conserved pore-forming region (known as H5 or SS1-SS2).Our results suggest that the structural motif for K+ channelshas been conserved between plants and animals.
68                                   NAL Call. No.: 381 AR2Functional expression of mammalian cytochromes P450IIB in theyeast 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; Genetictransformation; Cytochromes; Gene expression; Mammals
Abstract:  Three mammalian cytochromes P450 from the IIBsubfamily, P450IIB11 from canine and P450IIB4 and P450IIB5from rabbit, have been expressed in the yeast Saccharomycescerevisiae by use of an autonomously replicating vectorcontaining the galactose-inducible gal10 promoter. CytochromesP450IIB4 and P450IIB5 are closely related proteins, with only11 amino acid substitutions between them. P450IIB11 is ahomologous protein, likely orthologous with IIB4 or IIB5, with102 amino acid substitutions compared with the P450IIB4protein and 106 compared with the P450IIB5 protein. Theexpressed proteins are functional in yeast microsomes,exhibiting activity toward androstenedione, 7-ethoxycoumarin,and, in some cases, progesterone. Expressed cytochromesP450IIB4 and P450IIB11 hydroxylate androstenedione with regioand stereoselectivity characteristic of the purified,reconstituted proteins. A striking difference in theandrostenedione metabolite profiles of IIB4 and IIB5 wasobserved, with IIB4 producing almost exclusively the 16 betahydroxy metabolite and IIB5 producing the 16 alpha-hydroxy and15 alpha-hydroxy products. This is the first time that 15alpha-hydroxylase activity has been associated with IIB4/IIB5.This activity has also been detected in liver microsomes fromsome, but not all, individual phenobarbital-induced rabbitstested and is largely inhibited by anti-rabbit P450IIBimmunoglobulin G. These studies illustrate the utility of theyeast expression system for defining catalytic activities ofindividual mammalian cytochromes P450 and identifying newmarker activities that can be utilized in liver microsomes.
69                                  NAL Call. No.: 381 J824Functional expression of plant plasma membrane H+-ATPase inyeast endoplasmic reticulum.Villalba, J.M.; Palmgren, M.G.; Berberian, G.E.; Ferguson, C.;Serrano, R. Baltimore, Md. : American Society for Biochemistryand 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 hasbeen produced in a yeast expression system comprising amulticopy plasmid and the strong promoter of the yeast PMA1gene. Western blotting with a specific monoclonal antibodyshowed that the plant ATPase is one of the major membraneproteins made by the transformed cells, accounting for about1% of total yeast protein. The plant ATPase synthesized inyeast is fully active. It hydrolyzes ATP, pumps protons, andthe reaction cycle involves a phosphorylated intermediate.Phosphorylation is possible from both ATP and Pi. Unlike thesituation in plants, however, most of the plant ATPase is notexpressed in the yeast plasma membrane. Rather, the enzymeappears to remain trapped at a very early stage of secretorypathway: insertion into the endoplasmic reticulum. Thisorganelle was observed to proliferate in the form of stackedmembranes surrounding the yeast nucleus in order toaccommodate the large amount of plant ATPase produced. In thislocation, the plant ATPase can be purified with high yield (70mg from 1 kg of yeast) from membranes devoid of endogenousyeast plasma membrane H+-ATPase. This convenient expressionsystem could be useful for other eukaryotic membrane proteinsand ATPases.
70                               NAL Call. No.: SB732.6.M65The Fusarium solani-induced expression of a pea gene familyencoding 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 C162Gene expression associated with light-induced conidiation inColletotrichum 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 J824Half-calmodulin is sufficient for cell proliferation.Expressions of N-and C-terminal halves of calmodulin in theyeast Saccharomyces cerevisiae. Sun, G.H.; Ohya, Y.; Anraku,Y.Baltimore, Md. : American Society for Biochemistry andMolecular Biology; 1991 Apr15.The Journal of biological chemistry v. 266 (11): p. 7008-7015;1991 Apr15. Includes references.
Language:  English
Descriptors: Saccharomyces cerevisiae; Calmodulin; Geneexpression; Cell division; Structure activity relationships
Abstract:  Calmodulin (CaM) has been shown to be an essentialcomponent for progression of nuclear division in the yeastSaccharomyces cerevisiae (Ohya, Y., and Anraku, Y. (1989)Curr. Genet. 15, 113-120). To define the functional domain ofthe molecule required for cell proliferation, we constructedplasmids expressing a series of N- and C-terminal halves ofthe CaM under the control of the galactose-inducible GAL1promoter. These plasmids were introduced into a cmd1-disruptedyeast haploid strain, and the growth properties of the cellsdepending on the half-CaMs were examined. Plasmids expressingthe N-terminal half (Ser1-Leu176) and the C-terminal half(Leu85-Cys147), which each maintain two complete EF-handstructures, complemented the growth defect of the cmd1 nullmutation, whereas those expressing shorter regions of C- andN-terminal CaM did not. The half-CaMs that complemented thecmd1 null mutation were found to be approximately 6-foldoverexpressed relative to expression of native Cam by thewild-type CMD1 gene. The levels of expression of the half CaMswith the true CMD1 promoter were not sufficient forcomplementation. These results demonstrate that half-CaMs(either the N- or the C-terminal) are capable of supportinggrowth of yeast cells when they are suitably overproduced.Cells depending solely on half-cams all showed a temperaturesensitive growth phenotype, suggesting that half-CaMs cannotcarry out all the cellular functions of the complete CaMmolecule.
73                                   NAL Call. No.: 381 AR2Heterologous expression and characterization of an activelignin peroxidase from Phanerochaete chrysosporium usingrecombinant 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; Ligninolyticmicroorganisms; Peroxidases; Isoenzymes; Genetic engineering
Abstract:  The cDNA clone lambdaML-1 encoding one of theextracellular lignin peroxidases from the white rot fungus,Phanerochaete chrysosporium, was heterologously expressed inan active form using a recombinant baculovirus system. Theglycosylated extracellular form of the recombinant proteincontained the ferriprotoporphyrin IX moiety and was capable ofoxidizing both iodide and the model lignin compound, veratrylalcohol. In comparative peroxidase assays using guaiacol andMN(II), the recombinant lignin peroxidase did not appear to beMN(II) dependent. Sodium dodecyl sulfate-polyacrylamide gelelectrophoresis demonstrated that the heterologously expressedperoxidase had an apparent molecular weight similar to that ofthe native fungal isozyme H8. The elution profile of theactive recombinant enzyme derived by ion-exchangechromatography and immunoblot analysis using an anti-H8monoclonal antibody provided further evidence that thelambdaML-1 DNA encodes the lignin peroxidase H8.
74                               NAL Call. No.: 442.8 B5236Heterologous expression of active manganese peroxidase fromPhanerochaete chrysosporium using the baculovirus expressionsystem.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; Nuclearpolyhedrosis viruses; Recombination; Enzyme activity; Geneexpression; Purification; Characterization
Abstract:  The cDNA encoding Mn peroxidase isozyme H4 fromPhanerochaete chrysosporium was recombined into a baculovirusand heterologously expressed in Sf9 cells. The recombinant Mnperoxidase has the same molecular weight as the native enzymeas determined by SDS-PAGE and cross-reacts with a Mnperoxidase-specific antibody. The recombinant enzyme has aslightly lower pI than the native fungal isozyme H4 indicatingsome differences in post-translational modification. Phenolred, guaiacol, and vanillylacetone, substrates of the nativeMn peroxidase, are oxidized by the recombinant enzyme. All ofthe activities axe dependent on both Mn (II) and H2O2.
75                                  NAL Call. No.: 381 J824Heterologous expression of peptide hormone precursors in theyeast Saccharomyces cerevisiae. Evidence for a novelprohormone endoprotease with specificity for monobasic aminoacids.Bourbonnais, Y.; Danoff, A.; Thomas, D.Y.; Shields, D.Baltimore, Md. : American Society for Biochemistry andMolecular Biology; 1991 Jul15.The Journal of biological chemistry v. 266 (20): p.13203-13209; 1991 Jul15. Includes references.
Language:  English
Descriptors: Saccharomyces cerevisiae; Somatostatin; Geneexpression; Plasmids; Cloning; Precursors; Mammals
Abstract:  The peptide somatostatin (SRIF) exists as twodifferent molecular species. In addition to the most commonform, which is a 14-residue peptide, there is also a 14-aminoacid amino-terminally extended form of the tetradecapeptide,SRIF-28. Both peptides are synthesized as larger precursorscontaining paired basic and monobasic amino acids at theirprocessing sites, which, upon cleavage, generate eitherSRIF-14 or -28, respectively. In mammals a single prepro-SRIFmolecule undergoes tissue-specific processing to generate themature hormone whereas in some species of fish separate genesencode two distinct but homologous precursors prepro-SRIF-Iand -II that give rise to SRIF-14 and -28, respectively. Toinvestigate the molecular basis for differential processing ofthe prohormones we introduce their cDNAs into yeast cells(Saccharomyces cerevisiae). The signal peptides of bothprecursors were poorly recognized by the yeast endoplasmicreticulum translocation apparatus, consequently only lowlevels of SRIF peptides were synthesized. To circumvent thisproblem a chimeric precursor consisting of the alpha-factorsignal peptide plus 30 residues of the proregion was fused topro-SRIF-II. This fusion protein was efficiently transportedthrough the yeast secretory pathway and processed to SRIF-28exclusively, which is identical to the processing of thenative precursor in pancreatic islet D-cells. Mostsignificantly, cleavage of the precursor to SRIF-28 wasindependent of the Kex 2 endoprotease since processingoccurred efficiently in a kex 2 mutant strain. We concludethat in addition to the Kex 2 protease, yeast possess adistinct prohormone converting enzyme with specificity towardmonobasic processing sites.
76                                  NAL Call. No.: Q320.B56Heterologous 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 AR26High level expression of isocitrate lyase gene of n-alkaneutilizing yeast Candida tropicalis in Saccharomycescerevisiae.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, Candidatropicalis, was truncated to utilize the original open readingframe under the control of the GAL7 promoter and was expressedin Saccharomyces cerevisiae. The recombinant ICL wassynthesized as a functionally active enzyme with a specificactivity, similar to the enzyme purified from C. tropicalis,and was accounted for approximately 30% of the totalextractable proteins in the yeast cells. This recombinantenzyme was easily purified to homogeneity. N-terminal aminoacid sequence, molecular masses of native form and subunit.amino acid composition, peptide maps, and kinetic parametersof the recombinant ICL were essentially the same its those ofICL purified from C. tropicalis. From these facts, S.cerevisiae was suggested to be an excellent microorganism tohighly express the genes encoding peroxisomal proteins of C.tropicalis.
78                                  NAL Call. No.: QH426.C8The identification of 18 nuclear genes required for theexpression of the yeast mitochondrial gene encoding cytochromec 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.  Includesreferences.
Language:  English
Descriptors: Saccharomyces cerevisiae; Mitochondrial  DNA;Genes; Cytochrome-c oxidase; Gene expression; Geneticregulation; Nuclei; Nucleocytoplasmic interaction; Inducedmutations; Mutants; Messenger  RNA; Rna editing;Transcription; Mitochondrial genetics
Abstract:  Eighteen nuclear mutants of the yeast Saccharomycescerevisiae, each disturbed in the biosynthesis of themitochondrially encoded cytochrome c oxidase subunit 1 (cox1)and each representing a distinct complementation group, havebeen examined to identify the level at which COX1 expressionis affected. RNA blotting revealed that most have a defect inthe processing of COX1 precursor-mRNA; only a few aredefective in COX1 transcription and/or pre-mRNA stability. Inmost RNA-processing mutants, the absence of the COX1 messengerresults from a defect in the splicing of one or more COX1introns. In turn, this defect can be ascribed to a mutation ina nuclear gene which is either directly involved in splicingor else acts indirectly by impairing COX1 translation.
79                                  NAL Call. No.: 500 N21PIdentification of a tomato gene for the ethylene-formingenzyme 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 UnitedStates of America v. 88 (16): p. 7434-7437; 1991 Aug15. Includes references.
Language:  English
Descriptors: Lycopersicon esculentum; Saccharomycescerevisiae; Amino acid sequences; Biosynthesis; Ethylene;Enzyme activity; Gene expression; Genetic transformation;Molecular genetics; Nucleotide sequences; Oxidoreductases
Abstract:  The ethylene-forming enzyme (EFE), which catalyzesthe last step in the biosynthesis of the plant hormoneethylene, has never been purified and no molecular probes areavailable. Recently, a putative cDNA done for tomato EFE(pTOM13) has been identified by inhibiting ethylene synthesiswith an antisense gene expressed in transgenic plants. Adirect test of its function has been made by expression of apTOM13 gene in Saccharomyces cerevisiae. After cloningartefacts were discovered in the 5' region of the cDNA, acorrected cDNA (pRC13) was created by the fusion of the 5' endof a genomic clone to the 3' end of the cDNA and expressed inS. cerevisiae. Cultures of transformed yeast converted 1-aminocyclopropane-l-carboxylic acid (ACC) to ethylene, whereascontrol cells did not. This EFE activity displays similarcharacteristics to EFE found in plant tissue: it converts thetrans 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 theactivity of effectors on yeast EFE activity supports thehypothesis that EFE is one of a group of hydroxylase enzymes.
80                                  NAL Call. No.: 500 N21PIdentification of Dictyostelium Galpha genes expressed duringmulticellular 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 UnitedStates of America v. 88 (18): p. 8213-8217; 1991 Sep15. Includes references.
Language:  English
Descriptors: Dictyostelium; Amino acid sequences; Bindingproteins; Gene expression; Gene mapping; Genetic analysis;Genetic code; Guanine; Nucleotide sequences; Polymerase chainreaction
Abstract:  Guanine nucleotide-binding protein (G protein)-mediated signal transduction constitutes a common mechanism bywhich cells receive and respond to a diverse set ofenvironmental signals. Many of the signals involved in thedevelopmental life cycle of the slime mold Dictyostelium havebeen postulated to be transduced by such pathways and, in somecases, these pathways have been demonstrated to be dependenton 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 expressedduring the multicellular stages of development, suggestingpossible roles in cell differentiation or morphogenesis. Theentire G alpha 4 gene was sequenced and found to encode aprotein consisting of 345 amino acids. The G alpha 4 subunitis homologous to other previously identified G alpha subunits,including the Dictyostelium G alpha 1 (43% identity) and Galpha 2 (41% identity) subunits. However, the G alpha 4subunit contains some unusual sequence divergences in residueshighly conserved among most eukaryotic G alpha subunits,suggesting that G alpha 4 may be a member of another class ofG alpha subunits.
81                                NAL Call. No.: QH442.A1G4Improved 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; Geneexpression; Vectors
Abstract:  New yeast episomal vectors having a high degree ofutility for cloning and expression in Saccharomyces cerevisiaeare described. One vector, pYEULlacZ, is based on pUC19 andemploys the pUC19 multiple cloning site for the selection ofrecombinants in Escherichia coli by lacZ inactivation. Inaddition, the vector contains two genes, URA3 and leu2-d, forselection of the plasmid in ura3 or leu2 yeast strains. Thepresence of the leu2-d gene appears to promote replication athigh copy numbers. The introduction of CUP1 cassettes allowsthese plasmids to direct Cu2+-regulated production of foreignproteins in yeast. We show the production of a helminthantigen as an example of the vector application.
82                                  NAL Call. No.: 500 N21P Invitro assembly of apophytochrome and apophytochrome deletionmutants 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 UnitedStates 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 apoproteinexpressed in yeast is shown to assemble in vitro withphycocyanobilin to produce a photoreversible phytochrome-likeadduct. As an initial investigation of the amino acid sequencerequirements for chromophore incorporation, three phyA geneproduct deletion mutants were produced in yeast. Truncation ofthe N-terminal tail to residue 46 demonstrates that thisregion is not critical to bilin attachment, but a deletionmutant lacking 222 amino acids from the N terminus failed toyield holophytochrome in vitro, under the same conditions. Amutant comprising a deletion of the C terminus to residue 548showed bilin incorporation and red/far-red photoreversibility,indicating that bilin-apophytochrome assembly still occurredeven when the entire C-terminal domain was truncated.
83                                 NAL Call. No.: 442.8 Z34Inhibition of cell proliferation in Saccharomyces cerevisiaeby expression of human NAD+ ADP-ribosyltransferase requiresthe 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; Geneexpression; Dna; Binding site; Cell division; Chemicalreactions; 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 enzymein Saccharomyces cerevisiae, under the control of the alcoholdehydrogenase promoter, was only possible with simultaneousinhibition of ADP-ribosylation by 3-methoxybenzamide.Induction of fully active pADPRT from the inducible galactoseepimerase promoter resulted in inhibition of cell division andmorphological changes reminiscent of cell cycle mutants.Expression of a pADPRT cDNA truncated at its 5' end had noinfluence 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 pADPRTactivity by DNA breaks, is also required for inhibition ofcell growth on expression in yeast. Full-length as well astruncated pADPRT molecules were directed to the cell nucleuswhere the fully active enzyme produced large amounts ofpoly(ADP-ribose) by automodification. Since pADPRT turned outto be the only target for ADP-ribosylation in these cells,elevated levels of poly(ADP-ribose) were the most likely causeof inhibition of cell division, presumably resulting frominteraction with chromosomal proteins.
84                                NAL Call. No.: QH442.A1G4The K2-type killer toxin- and immunity-encoding region fromSaccharomyces 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 acidsequences; Plant proteins; Genetic code; Resistance
Abstract:  The cDNA copies of M2-1, the larger heat-cleavageproduct of M2 double-stranded (ds) RNA, have been synthesized,cloned, sequenced and expressed in yeast. This sequence, incombination with the known terminal sequence of M2-1 dsRNA,identifies a translation reading frame for a 362-amino-acidprotein of 38.7 kDa, similar in size to the one of severalprotein species produced from M2-1 dsRNA in vitro translation.The expression of this cDNA clone in yeast confers both killerand immunity phenotypes.
85                                 NAL Call. No.: 448.3 AP5Manganese regulation of manganese peroxidase expression andlignin degradation by the white rot fungus Dichomitussqualens.Perie, F.H.; Gold, M.H.Washington, D.C. : American Society for Microbiology; 1991Aug. 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 laccaseactivities were detected in cultures of Dichomitus squalens(Polyporus anceps) under conditions favoring lignindegradation. In contrast, neither extracellular ligninperoxidase nor aryl alcohol oxidase activity was detected incultures grown under a wide variety of conditions. Themineralization of 14C-ring-, -side chain-, and -methoxylabeled synthetic guaiacyl lignins by D. squalens and theexpression of extracellular manganese peroxidase weredependent on the presence of Mn(II), suggesting that manganeseperoxidase is an important component or this organism's lignindegradation system. The expression of laccase activity wasindependent of manganese. In contrast to previous findingswith Phanerochaete chrysosporium, lignin degradation by D.squalens proceeded in the cultures containing excess carbonand nitrogen.
86                                  NAL Call. No.: QH426.C8The mating type in fission yeast is switched independently ofits expression. Ruusala, T.Berlin, W. Ger. : Springer International; 1991.Current genetics v. 20 (5): p. 379-383; 1991.  Includesreferences.
Language:  English
Descriptors: Endomycetales; Loci; Mating; Genetic change; Geneexpression; Transcription; Transposable elements; Plasmids
Abstract:  The mating type of fission yeast is determined bythe mat1 locus on chromosome II. The sequence content of thislocus, and hence the mating type, is switched in a strictlyregular pattern by transposition from one of two unexpressedmating type sequences. The expressed and the two silentsequences are located on the same chromosome. It is notunderstood how one of the two donor sequences is selected inthis reaction. Here I test the possibility that the selectionis governed by gene expression from the mat1 locus. Such amechanism could favor transposition of a donor sequence ofopposite mating type to the one present at mat1. Alternativelyit could disfavor transposition of a synonymous sequence. Thepresent data argue strongly against any type of participationof mat1 gene products in the choice of donor during the matingtype switch. Alternative steering mechanisms are discussed.
87                                  NAL Call. No.: QR53.B56Methanol 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 ofmethanol to yeast cultures was found to give strong inductionof a lacZ gene under heat shock element (HSE) control.Methanol addition is therefore an easily-executed alternativeto temperature upshift for the induction of HSE-directed geneexpression, although in this study it gave slightly lowerlevels of induction compared to heat shock.
88                                 NAL Call. No.: QK600.B72Methylotrophic 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 moleculargenetics of fungi / edited by J. F. Peberdy, C. E. Caten, J.E. Ogden and J. W. Bennett. Symposium of the BritishMycological Society held at the University of Nottingham,April 1990. Literature review.  Includes references.
Language:  English
Descriptors: Pichia; Hansenula; Genetic transformation; Genetransfer; Gene expression; Structural genes; Proteins;Plasmids; Vectors; Alcohol oxidoreductases; Promoters;Methanol; Substrates; Literature reviews
89                                 NAL Call. No.: QP501.B64Mitochondrial gene expression in Saccharomyces cerevisiae:proteolysis of nascent chains in isolated yeast mitochondriaoptimized for protein synthesis. Black-Schaefer, L.; McCourt,J.D.; Poyton, R.O.; McKee, E.E. London : The BiochemicalSociety; 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; Proteinsynthesis; Translation; Proteolysis; Gene expression
Abstract:  We demonstrate here that mitochondrial translationproducts synthesized by isolated yeast mitochondria aresubject to rapid proteolysis. The loss of label frommitochondrial peptides synthesized in vitro comes from twodistinct pools of peptides: one that is rapidly degraded (t1/2of minutes) and one that is much more resistant to proteolysis(t1/2 of hours). As the length of the incubation periodincreases, the percentage of labelled peptides in the rapidlyturning-over pool decreases and cannot be detected after 60min of incubation. This proteolysis is inhibited bychloramphenicol and is dependent on the presence of ATP. Theloss of label during the chase occurs from fully completedtranslation products. The proteolysis observed here markedlyaffects measurements of rates of mitochondrial proteinsynthesis in isolated yeast mitochondria. In earlier work, inwhich proteolysis was not considered, mitochondrialtranslation was thought to stop after 20-30 min of incubation.In the present study, by taking proteolysis into account, wedemonstrate that the rate of translation in isolatedmitochondria is actually constant for nearly 60 min and thendecreases to near zero by 80 min of incorporation. Thesefindings have allowed us to devise a procedure for measuringthe 'true' rate of translation in isolated mitochondria. Inaddition, they suggest that mitochondrial translation productswhich normally assemble with nuclear-encoded gene productsinto multimeric enzyme complexes are unstable without theirnuclear-encoded counterparts.
90                             NAL Call. No.: QK1.D5 Bd.166Mitochondriale Genexpression bei Pilzen molekulare Analysenzur nukleo-zytoplasmatischen Wechselwirkung  [Mitochondrialgene expression in fungi].Heinen, UteBerlin : 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.M65Molecular analysis of the laccase gene from the chestnutblight fungus and selective suppression of its expression inan 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; Plantpathogenic fungi; Nucleotide sequences; Amino acid sequences;Comparisons; Neurospora crassa; Laccase; Genes; Speciesdifferences; Strain differences; Hypovirulence; Strains;Messenger  RNA; Tubulin; Actin; Glyceraldehyde-3-phosphatedehydrogenase; Gene expression; Genetic analysis; Introns
92                                  NAL Call. No.: QK600.S4The molecular biology of Trichoderma and its application tothe 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 seriesanalytic: Molecular industrial mycology: systems andapplications for filamentous fungi / edited by S. A. Leong andR. 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 N21PMolecular cloning and expression in photosynthetic bacteria ofa soybean cDNA coding for phytoene desaturase, an enzyme ofthe 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 UnitedStates 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 redphotoprotective pigments present in all plastids. The firstcarotenoid of the pathway is phytoene, a colorless compoundthat is converted into colored carotenoids through a series ofdesaturation reactions. Genes coding for carotenoiddesaturases have been cloned from microbes but not fromplants. We report the cloning of a cDNA for pds1, a soybean(Glycine max) gene that, based on a complementation assayusing the photosynthetic bacterium Rhodobacter capsulatus,codes for an enzymic that catalyzes the two desaturationreactions that convert phytoene into zeta-carotene, a yellow,carotenoid. The 2281-base-pair cDNA clone analyzed contains anopen reading frame with the capacity to code for a 572-residueprotein of predicted Mr 63,851. Alignment of the deduced Pds1peptide sequence with the sequences of fungal and bacterialcarotenoid desaturases revealed conservation of several aminoacid residues, including a dinucleotide-binding motif thatcould mediate binding to FAD. The Pds1 protein is synthesizedin vitro as a precursor that, upon import into isolatedchloroplasts, is processed to a smaller mature form.Hybridization of the pds1 cDNA to genomic blots indicated thatthis gene is a member of a low-copy-number gene family. One ofthem loci was genetically mapped using restriction fragmentlength polymorphisms between Glycine max and Glycine soja. Weconclude that pds1 is a nuclear gene encoding a phytoenedesaturase enzyme that, as its microbial counterparts,contains sequence motifs characteristic of flavoproteins.
94                                  NAL Call. No.: 500 N21PMolecular cloning, expression, and induction of berberinebridge enzyme, an enzyme essential to the formation ofbenzophenanthridine alkaloids in the response of plants topathogenic attack.Dittrich, H.; Kutchan, T.M.Washington, D.C. : The Academy; 1991 Nov15.Proceedings of the National Academy of Sciences of the UnitedStates of America v. 88 (22): p. 9969-9973; 1991 Nov15. Includes references.
Language:  English
Descriptors: Eschscholzia californica; Cloning; Geneexpression; Molecular genetics; Oxidoreductases; Phytoalexins;Biosynthesis; Enzyme activity; Nucleotide sequences;Saccharomyces cerevisiae; Amino acid sequences; Fungaldiseases
Abstract:  The berberine bridge enzyme [(S)-reticuline: oxygenoxidoreductase (methylene-bridge-forming), EC 1.5.3.9] is avesicular plant enzyme that catalyzes the formation of theberberine bridgehead carbon of (S)-scoulerine from the Nmethyl carbon of (S)-reticuline in a specific, unparalleledreaction along the biosynthetic pathway that leads tobenzophenanthridine alkaloids. Cytotoxic benzophenanthridinealkaloids are accumulated in certain species of Papaveraceaeand Fumariaceae in response to pathogenic attack and,therefore, function as phytoalexins. The berberine bridgeenzyme has been purified to homogeneity from elicited cellsuspension cultures of Eschscholtzia californica, and partialamino acid sequences have been determined. A cDNA, isolatedfrom a lambda-gt11 cDNA bank of elicited E. californica cellsuspension cultures, coded for an open reading frame of 538amino acids. The first 22 amino acids constitute the putativesignal peptide. The mature protein has a Mr of 57,352,excluding carbohydrate. The berberine bridge enzyme washeterologously expressed in a catalytically active form inSaccharomyces cerevisiae. Southern hybridization with genomicDNA suggests that there is only one gene for the enzyme in theE. californica genome. Hybridized RNA blots from elicited E.californica cell-suspension cultures revealed a rapid andtransient increase in poly(A)+ RNA levels that preceded boththe increase in enzyme activity and the accumulation ofbenzophenanthridine alkaloids, emphasizing the integral roleof the berberine bridge enzyme in the plant response topathogens.
95                                   NAL Call. No.: QR1.F44Molecular cloning of genes from the rumen anaerobic fungusNeocallimastix frontalis: expression during hydrolaseinduction.Reymond, P.; Durand, R.; Hebraud, M.; Fevre, M.Amsterdam : Elsevier Science Publishers; 1991 Jan01.FEMS microbiology letters - Federation of EuropeanMicrobiological Societies v. 77 (1): p. 107-112; 1991 Jan01. Includes references.
Language:  English
Descriptors: Neocallimastix; Rumen fungi; Anaerobes;Hydrolases; Induction; Dna hybridization; Rna; Cloning; Geneexpression; Screening; Dna libraries; Translation;Transcription; Nucleotide sequences
Abstract:  Glycoside and polysaccharide hydrolase productionby the rumen anaerobic fungus, Neocallimastix frontalis isinduced by the presence of crystalline cellulose. Adifferential screening of a cDNA library was used to isolateDNA sequences transcribed at high levels under growthconditions which induce enzyme production. Seven clones wereisolated that prefentially hybridized to the induced cDNAprobe versus the non-induced cDNA probe. Southern analysisshowed that a cDNA clone (118) hybridized to a DNA probeencoding part of the exo-cellobiohydrolase I (CBH I) gene ofTrichoderma reesei. Northern analysis demonstrated that thecDNA 118 was transcribed to yield a 2.1 kb RNA. Thistranscript was induced in the presence of cellulose.
96                                  NAL Call. No.: 500 N21PMonitoring of intracellular calcium in Saccharomycescerevisiae 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 UnitedStates of America v. 88 (15): p. 6878-6882; 1991 Aug01. Includes references.
Language:  English
Descriptors: Saccharomyces cerevisiae; Calcium ions; Cellgrowth; Gene expression; Luminescence; Plant proteins;Recombinant  DNA; Sex pheromones
Abstract:  A method is described for measuring cytosolic freeCa2+ and its time-dependent changes in the yeast Saccharomycescerevisiae by using the luminescent protein aequorin as aCa2+-specific indicator. This method with intact yeast cellsis labeled "in vivo" to distinguish it from methods with cellextracts, labeled "in vitro." A plasmid in which theapoaequorin cDNA was joined downstream from theglyceraldehyde-3-phosphate dehydrogenase gene promoter wasconstructed and introduced into yeast cells. The intracellularconcentration of apoaequorin expressed by the cDNA wasapproximately 1 micromole, which was high enough to detect thecytosolic Ca2+. Growth of the transformed cells was normal. Inthe in vitro method, apoaequorin in crude cell extracts wasregenerated into aequorin by mixing with coelenterazine, thesubstrate for the luminescence reaction, whereas in the invivo method, aequorin was regenerated by incubating intactcells with coelenterazine. Simultaneous addition of 10 mMCaCl2 and 10 micromole A23187, a Ca2+ ionophore, tocoelenterazine-incorporated cells generated luminescence.Coelenterazine-incorporated cells also responded to nativeextracellular stimuli. A mating pheromone, alpha-factor, addedto cells of mating type a or alpha, generated extracellularCa2+-dependent luminescence specifically in a mating typecells, with maximal intensity occurring 45-50 min afteraddition of alpha-factor. Glucose added to glucose-starvedG0/G1 cells stimulated an increase in extracellular Ca2+-dependent luminescence with maximal intensity occurring 2 minafter addition. These results show the usefulness of theaequorin system in monitoring [Ca2+]i, response toextracellular stimuli in yeast cells.
97                                  NAL Call. No.: 381 J824Multiple regulatory elements control expression of the geneencoding the Saccharomyces cerevisiae cytochrome P450,lanosterol 14 alpha-demethylase (ERG11).Turi, T.G.; Loper, J.C.Baltimore, Md. : American Society for Biochemistry andMolecular 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; Geneexpression; Genetic regulation
Abstract:  The major cytochrome P450 in the yeastSaccharomyces cerevisiae, lanosterol 14 alpha-demethylase(ERG11), catalyzes an essential reaction in the biosynthesisof ergosterol, the predominant sterol of yeast. Protein levelsof this cytochrome P450 are known to be affected by carbonsource, oxygen, and heme, as well as the growth state of theculture. We have determined that ERG11 message levels increaseduring growth on glucose, in the presence of heme, and duringoxygen limiting growth conditions and, unexpectedly, duringanaerobic growth. To determine the cis-acting regionsresponsible for regulation of expression of the ERG11 promoterunder optimal conditions of fermentative growth, deletionanalysis was performed using the Escherichia coli lacZ as areporter gene. Two upstream activating sequences, UAS1 andUAS2, and an upstream repressor element, URS1, plus a secondpossible or cryptic repressor element, URS2, were identifiedin the ERG11 promoter. The HAP1 protein product apparentlyparticipates in activation from UAS1 but not from UAS2.Sequences resembling ERG11 UAS2 were identified in sevenadditional oxygen-regulated genes. Repression of ERG11,expression was dependent upon the ROX1 repressor andadditional repressor(s) designated as Old (overexpression oflanosterol demethylase). These data indicate that ERG11 is amember 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 becoordinately regulated with ERG11.
98                                NAL Call. No.: QH442.A1G4 Anovel, highly regulated, rapidly inducible system for theexpression of chicken progesterone receptor, cPRA, inSaccharomyces 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; Hormonereceptors; Progesterone; Genes; Fowls; Gene expression;Transcription; Controlling elements
Abstract:  A rapidly inducible and tightly regulated systemfor the expression of protein in yeast is based on a chimericpromoter constructed of two copies of a vitellogenin-estrogenresponse element (ERE) which are inserted upstream from thepromoter of the yeast gene encoding iso-1-cytochrome c. Thechimeric promoter was inserted in a yeast expression plasmidupstream from the coding sequence of ubiquitin fused in frameto a cDNA encoding the full-length chicken progesteronereceptor A (cPR(A)). The resultant plasmid (YEpA2) was cotransformed in Saccharomyces cerevisiae with a plasmid whichencodes the human estrogen receptor. Estradiol (E2)-inducedtransactivation of the chimeric promoter results intranscription of the cPR(A) gene from YEpA2 and synthesis ofcPR(A). The fusion protein, ubiquitin-cPR(A), is rapidlycleaved in vivo to produce cPR(A). Analysis of samples byWestern immunoblot shows that cPR(A) is almost undetectable inthe absence of E2, and that treatment with 50 nM E2 results ina 500-1000-fold induction of cPR(A) (0.06-0.3% of the totalprotein) after 1 h. The plasmid-expressed soluble receptor isstable and demonstrates the correct affinity for its ligand.We have prepared yeast extracts using enzymatic digestion ofthe cell wall with oxalyticase followed by hypotonic shock.This has resulted in a dramatic increase in the % of receptorwhich binds hormone compared to previous studies which usedmechanical disruption techniques. The cPR(A) is biologicallyactive since it activates transcription of a co-transformedreporter gene containing its response element. This system iswell suited for the regulated synthesis of proteins in yeastand may be especially useful for producing labile proteins orproteins toxic to the cells.
99                               NAL Call. No.: QH431.A1G43Nuclear-cytoplasmic interactions in the resistance of wheat tofungal pathogens. II. Effects of cultivated and wild cerealcytoplasms on the expression of the genome of the Leningradkavariety 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; Aegilopsovata; Triticum spelta; Plant pathogenic fungi; Plantbreeding; Substitution lines; Genetic resistance; Mildews;Fungal diseases; Nucleocytoplasmic interaction; Cytoplasmicinheritance; Plant development; Crop growth stage; Sporulation
Abstract:  The quantitative resistance of three alloplasmaticand 29 substituted soft wheat lines containing the nucleargenome of the Leningradka variety to a floury mildewpopulation was studied on a stringent infective background, bymeasuring the mean spore-bearing ability of the fungus per cm2of the third and flag leaves of each plant line and theinitial Leningradka variety. Dispersion analysis showed thatthe plant cytoplasm had a significant effect on pathogenreproduction during the ontogenetic development of wheatplants. Cytoplasms from a number of cultivated and wildspecies had modifying effects on the expression of the nucleargenes controlling quantitative resistance. It is suggestedthat a specific interaction between the fungal and host plantgenomes is involved in determining quantitative resistance inthe presence of cytoplasms modifying fungal reproductionduring plant ontogenesis. A non-specific interaction probablyoccurs in the presence of cytoplasms maintaining a stablelevel of quantitative resistance during ontogenesis.
100                                 NAL Call. No.: QH426.C8Optimisation of a host/vector system for heterologous geneexpression 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.  Includesreferences.
Language:  English
Descriptors: Endomycetales; Vectors; Plasmids; Gene transfer;Gene expression; Cyamopsis tetragonoloba; Alpha-galactosidase;Reporter genes; Saccharomyces cerevisiae; Betafructofuranosidase; Genetic transformation; Nucleotidesequences; Oxidoreductases
Abstract:  For the methylotrophic yeast, Hansenula polymorpha,expression vectors with different origins of replication havebeen constructed in order to analyse their influence ontransformation and integration efficiency. The constructedplasmids are identical except for their origin of replication,which involve, respectively, that of the Saccharomycescerevisiae 2-mu m plasmid and a H. polymorpha ARS sequence(HARS2). A plasmid with no origin of replication served as acontrol. The plasmids also contained the alpha-galactosidaseexpression cassette, consisting of the Cyamopsis tetragonolobaalpha-galactosidase gene, the H. polymorpha methanol oxidasepromoter and terminator, and the S. cerevisiae invertasesignal sequence. The transformation frequencies of theexpression vectors containing the 2-mu m and the HARS2 originsof replication, and no origin of replication, were 2, 50 and15 per microgram of DNA respectively, which demonstrates thenegative effect of the 2-mu m sequence on the transformationfrequency. Autonomously replicating plasmids could be isolatedfrom the transformants obtained with the plasmid containingeither the 2-mu m or the HARS2 sequence. Integration of the 2-mu m based plasmid into the H. polymorpha genome could not beestablished using a standard procedure. This is in contrastwith transformants containing a plasmid bearing the HARS2sequence or else with no origin of replication, which showsthat the 2-mu m sequence negatively influences the integrationof the expression vector into the H. polymorpha genome.Integration of expression plasmids occurred in 50% of theanalysed integrants on the homologous methanol oxidase locus,and tandem integration was favoured. The level of specificmRNA, and the expression of the alpha-galactosidase protein bythese integrants, was proportional to the number of integratedcopies of the expression plasmid in the H. polymorpha genome.
101                                NAL Call. No.: 442.8 Z34The organization and expression of the Saccharomycescerevisiae L4 ribosomal protein genes and their identificationas 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 twohomologous genes from Saccharomyces cerevisiae. The two yeastgenes (L4-1 and L4-2) were identified as encoding S.cerevisiae L4 by 2D gel analysis of the product of the invitro translation of hybrid-selected mRNA and additionally bydirect amino acid sequencing. The DNA sequences of the twoyeast genes were highly homologous (95%) over the 771 bp thatencode the 256 amino acids of the coding regions but showedlittle homology outside the coding region. L4-1 differed fromL4-2 by 7 out of the 256 amino acids in the coding region,which is the greatest divergence between the products of anytwo duplicated yeast ribosomal protein genes so far reported.There is strong homology between the mouse rpL7a/Surf-3 andthe yeast L4 genes -57% at the nucleic acid level and also 57%at the amino acid level (though some regions reach as much as80-90% homology). While most yeast ribosomal protein genescontain an intron in their 5' region both L4-1 and L4-2 areintronless. The mRNAs derived from each yeast gene containedheterogenous 5' and 3' ends but in each case the untranslatedleaders were short. The L4-1 mRNA was found to be much moreabundant than the L4-2 mRNA as assessed by cDNA andtranscription analyses. Yeast cells containing a disruption ofthe L4-1 gene formed much smaller colonies than either wildtype or disrupted L4-2 strains. Disruption of both L4 genes isa lethal event, probably due to an inability to producefunctional ribosomes.
102                                 NAL Call. No.: QH426.C8Over-expression, purification and determination of theproteolytic processing site of the yeast mitochonrial CBS1protein.Korte, A.; Michaelis, U.; Lottspeich, F.; Rodel, G.Berlin, W. Ger. : Springer International; 1991.Current genetics v. 20 (1/2): p. 87-90; 1991.  Includesreferences.
Language:  English
Descriptors: Saccharomyces cerevisiae; Mitochondria; Plantproteins; Purification; Amino acid sequences; Proteolysis;Genes; Genetic transformation; Gene expression
Abstract:  Yeast transformants harboring the CBS1 gene underthe control of the strong ADC1 promoter on a high copy numberplasmid express the mitochondrial CBS1 protein at artificiallyhigh levels. Over-expressed protein is imported intomitochondria and correctly processed to yield the maturemitochondrial 23.5 kDa form, but differs in its solubilityproperties from CBS1 in wild-type mitochondria. It formsinsoluble protein aggregates, which are refractory tosolubilization with 1% Taurodeoxycholate. We exploited thisobservation to separate CBS1 from the bulk of mitochondrialproteins and to isolate CBS1 after SDS gel electrophoresis.Determination of the amino-terminal amino acids of thepurified protein reveals that the mature CBS1 protein startswith Ile(30), at the characteristic distance of +2 amino acidsfrom an arginine residue (Arg(28)). The cleavage site shows aremarkable homology to that of subunit 9 of the F0F1 ATPasefrom Neurospora crassa.
103                                 NAL Call. No.: QH426.C8PDC6, a weakly expressed pyruvate decarboxylase gene fromyeast, is activated when fused spontaneously under the controlof the PDC1 promoter. Hohmann, S.Berlin, W. Ger. : Springer International; 1991.Current genetics v. 20 (5): p. 373-378; 1991.  Includesreferences.
Language:  English
Descriptors: Saccharomyces cerevisiae; Structural genes;Pyruvate decarboxylase; Alleles; Mutants; Promoters; Geneexpression; Genetic regulation; Genetic engineering; Enzymeactivity; Restriction mapping; Genetic code; Nucleotidesequences; Amino acid sequences
Abstract:  Three structural genes encode the pyruvatedecarboxylase isoenzymes in the yeast Saccharomycescerevisiae. PDC1 and PDC5 are active during glucosefermentation where PDC1 is expressed about six times morestrongly than PDC5. Expression of PDC6 is weak and seems to beinduced in ethanol medium. Consequently, pdc1 delta pdc5 deltadouble mutants do not ferment glucose and do not grow onglucose medium. Spontaneous mutants, derived from such a pdc1pdc5 strain, were isolated which could again ferment glucose.They showed pyruvate decarboxylase activity due to aduplication of PDC6. The second copy of PDC6 was expressedunder the control of the PDC1 promoter, which was stillpresent in the pdc1 strain. However, the resulting PDC1-PDC6fusion gene could only partially substitute for PDC1: toachieve normal growth and high pyruvate decarboxylase activitystrains carrying PDC1-PDC6 required a functional PDC5 genewhich is dispensable in a PDC1 wild-type background. Thus,expression of PDC5 depends on the state of the PDC1 locus: lowin the PDC1 wild-type background and high in PDC1-PDC6 fusionstrains and, as shown previously, in pdc1 mutants. Theactivation of PDC5 expression in PDC1-PDC6 strains may be dueto particular properties of the PDC1-PDC6 fusion protein orsimply to the weaker expression of PDC1-PDC6 in comparison tothe wild-type PDC1 gene.
104                                 NAL Call. No.: QP501.E8Phenylalanine amonia-lyase in potato (Solanum tuberosum L.).Genomic complexity, structural comparison of two selectedgenes 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; 1992Mar.  Includes references.
Language:  English
Descriptors: Solanum tuberosum; Phenylalanine ammonia-lyase;Structural genes; Messenger  RNA; Dna; Clones; Genomes; Aminoacid sequences; Nucleotide sequences; Phytophthora infestans;Injuries; Stress response; Gene expression
Abstract:  Potato (Solanum tuberosum L. cv. Datura) containsapproximately 40-50 phenylalanine ammonia-lyase (PAL)genes/haploid genome. Considerable cDNA heterogeneityindicates that at least about 10, and probably more, of thesegenes are potentially active. One subfamily, represented byone selected member (PAL-1), was analyzed with respect togenomic complexity, nucleotide and deduced amino acidsequence, and mode of constitutive or induced expression. Forcomparison, a second gene (PAL-2), representing severalsubfamilies that are easily distinguished from PAL-1, wasincluded in these studies. Extensive structural similaritieswere observed both between the TATA-proximal portions of thePAL-1 and PAL-2 promoters, particularly in the areascontaining putative cis-acting elements, and among allpresently known PAL proteins from various higher and lowerplants. The relative abundance of PAL mRNA varied greatly inseveral major potato organs. However, the patterns obtainedwith probes detecting either total PAL mRNA or morespecifically, PAL-1-related or PAL-2-related mRNA species,were the same within experimental error. Mature leavescontained particularly low levels of PAL mRNA. Infection ofthese leaves with the pathogenic fungus, Phytophthorainfestans, resulted in a large, transient induction of PALmRNA. The relative timing of PAL-1 and PAL-2 mRNA expression,however, differed in compatible (fungus virulent, plantsusceptible) but not in incompatible interactions (fungusavirulent, plant resistant). Wounding of leaves caused anextremely rapid and transient induction of both PAL mRNAspecies.
105                                NAL Call. No.: 448.3 J82Phosphatidylglycerolphosphate synthase expression inSchizosaccharomyces pombe is regulated by the phospholipidprecursors inositol and choline. Karkhoff-Schweizer, R.R.;Kelly, B.L.; Greenberg, M.L.Washington, D.C. : American Society for Microbiology; 1991Oct. Journal of bacteriology v. 173 (19): p. 6132-6138; 1991Oct.  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 committedstep in the cardiolipin biosynthetic pathway. To study theregulation of PGPS in Schizosaccharomyces pombe, wecharacterized the enzyme biochemically. Maximum activityoccurred in the presence of 6 mM Triton X-100 at pH 7.5. Theapparent Km values for CDP-diacylglycerol and glycerol 3-phosphate were 130 and 26 micromole, respectively. Optimalactivity was at 35 degree C, and enzyme activity was labileabove 40 degree C. Thioreactive agents were inhibitory to PGPSactivity. To determine whether S. pombe PGPS is regulated byphospholipid precursors, we examined the time-dependentexpression of PGPS upon inositol and choline starvation.Starvation for inositol resulted in a threefold increase inPGPS expression in wild-type cells. In cho1 and cho2 mutants,which are blocked in phosphatidylcholine synthesis, starvationfor choline resulted in transient derepression of PGPSexpression. In choline auxotrophs starved for inositol, PGPSwas derepressed 2.5- to 3-fold in the presence of choline andless or not at all in the absence of choline. This is thefirst description of PGPS regulation in S. pombe and the firstdemonstration of inositol-mediated regulation in the inositolrequiring yeast species.
106                               NAL Call. No.: QD341.A2N8Plant nonsense suppressor tRNATyr gene are expressed at verylow 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 togenerate amber, ochre and opal suppressors from clonedArabidopsis and Nicotiana tRNA(Tyr) genes. The nonsensesuppressor tRNA(Tyr) genes were efficiently transcribed inHeLa and yeast nuclear extracts, however, intron excision fromall mutant pre-tRNAsTyr was severely impaired in thehomologous wheat germ extract as well as in the yeast in vitrosplicing system. The change of one nucleotide in the anticodonof suppressor pre-tRNAs leads to a distortion of the potentialintron-anticodon interaction. In order to demonstrate thatthis caused the reduced splicing efficiency, we created apoint mutation in the intron of Arabidopsis tRNA(Tyr) whichaffected the interaction with the wild-type anticodon. Asexpected, the resulting pre-tRNA was also inefficientlyspliced. Another mutation in the intron, which restored thebase-pairing between the amber anticodon and the intron ofpre-tRNA(Tyr) resulted in an excellent substrate for wheatgerm splicing endonuclease. This type of amber suppressortRNA(Tyr) gene which yields high levels of mature tRNA(Tyr)should be useful for studying suppression in higher plants.
107                                 NAL Call. No.: 381 B522Posttranscriptional regulation of the expression of MET2 geneof Saccharomyces cerevisiae.Forlani, N.; Martegani, E.; Alberghina, L.Amsterdam : Elsevier Science Publishers; 1991 May02.Biochimica et biophysica acta : International journal ofbiochemistry and biophysics v. 1089 (1): p. 47-53; 1991 May02. Includes references.
Language:  English
Descriptors: Saccharomyces cerevisiae; Methionine;Biosynthesis; Genetic regulation; Amino acid metabolism; Geneexpression
Abstract:  The first step of the specific pathway formethionine biosynthesis in the yeast Saccharomyces cerevisiaeis catalyzed by the enzyme L-homoserine-O-acetyltransferase(HSTase) (EC 2.3.1.31), encoded by the MET2 gene. In order toascertain whether there is a posttranscriptional control onthe MET2 gene expression, as suggested by previous results onthe expression of the cloned gene, systems for high inducibleexpression of MET2 gene were constructed. In these constructsthe MET2 gene was cloned in yeast expression vectors under thecontrol of an inducible yeast GAL promoter element so that theMET2 was transcribed at very high levels under inducedconditions. Measurements or the specific mRNA levels showed astrong stimulation of MET2 gene transcription in yeasttransformants grown on galactose as carbon source,corresponding to 50-100-fold the repressed conditions, whileonly a 2-fold increase of the enzymatic activity was observed.In addition, no evidence of a strong induced polypeptide ofappropriate size on two dimensional gel electrophoresis wasobtained. To understand the functional role of the non-coding5' region of MET2 mRNA, we performed either a partial and acomplete deletion of the 5' leader sequence, but even withthese constructs an elevated mRNA level was not associated toa marked increase of the HSTase activity. These data supportthe idea of a posttranscriptional regulation of MET2 geneexpression and show that the untranslated region of thespecific mRNA is not involved in this regulatory mechanism.
108                                NAL Call. No.: QP601.M49Probing molecular structure and structural changes of voltagegated channel by expressing mutant channels in yeast andreconstituting 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 seriesanalytic: 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 J8223Properties and distribution of soybean proglycinin expressedin 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 ofsoybean. The high-level expression system of preproglycinincDNA in yeast was established. The signal sequence of theexpressed protein was cleaved at the correct site. However,most of the expressed proteins were insoluble due to theirinteraction with intracellular components at the acidicpolypeptide region. The expressed proteins separated from theintracellular components by ion-exchange chromatography in thepresence of urea were soluble without urea and self-assembledinto trimers. The insolubility of the expressed proteinscaused an accumulation of modified proteins with disturbedfolding. Immunocytochemical distribution demonstrated that theexpressed protein from the cDNA encoding preproglycininaccumulated in Golgi-like structure and organellas which maybe derived from Golgi-like structure, and one from the cDNAencoding proglycinin homologue protein was found in thecytosol.
110                                 NAL Call. No.: QH426.C8Properties of two nuclear pet mutants affecting expression ofthe 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.  Includesreferences.
Language:  English
Descriptors: Saccharomyces cerevisiae; Mutants; Mitochondria;Gene expression; Atp; Ligases; Temperature; Geneticregulation; Transcription
Abstract:  This study details the characteristics of twotemperature-conditional pet mutants of yeast, strains ts1860and ts379, which at the non-permissive temperature showdeficiencies in the formation of three mitochondrially encodedsubunits of the ATP synthase complex. By analysis ofmitochondrial translation products, and of mitochondrialtranscription in temperature shift experiments from thepermissive (22 degrees C) to the non-permissive (36 degrees C)temperature, it was concluded that the nuclear mutations inboth mutants primarily inhibit synthesis of ATP synthasesubunit 9, and that reductions in subunit 8 and 6 synthesisare secondary pleiotropic effects. Following transfer to 36degree C, cells of mutant ts379 display a near completeinhibition of subunit 9 synthesis within 1 h, coincident witha marked reduction in the level of the cognate oli1 mRNA. Onthe other hand, near complete inhibition of subunit 9synthesis in strain ts1860 occurs after 3 h at 36 degrees C,at which time there is little change in the level of subunit 9mRNA. In both mutants the mRNA levels for subunits 6 and 8 arenot significantly affected at the time of inhibition ofsubunit 9 synthesis. Provision of an alternative source ofsubunit 8, translated extra-mitochondrially for import intothe organelle, does not overcome the mutant phenotype ofeither mutant at 36 degrees C, confirming that subunit 8 isnot the sole or primary deficiency in each mutant. The mutantsindicate that the products of a least two nuclear genes(designated AEP1 and AEP2) are required for the expression ofthe mitochondrial oli1 gene and the synthesis of subunit 9.The product of the AEP1 gene (defective in mutant tsl860) isrequired for translation of oli1 mRNA while the AEP2 product(defective in mutant ts379) is essential either for thestability of oli1 mRNA or for the correct processing ofprecursor transcripts to the mature message.
111                                 NAL Call. No.: QR53.B56Proteolytic degradation of heterologous proteins expressed inAspergillus 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 proteinsexpressed in the filamentous fungus Aspergillus niger reducesthe yield of authentic target protein. The activities of A.niger proteases are differentiated by their effects on twoproteins expressed and secreted from A. niger: hen egg-whitelysozyme and porcine pancreatic phospholipase A2.
112                                 NAL Call. No.: QP501.E8Purification, characterisation and mutagenesis of highlyexpressed 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; 1991Jun.  Includes references.
Language:  English
Descriptors: Saccharomyces cerevisiae; Pyruvate kinase;Recombination; Gene expression; Purification;Characterization; Mutagenesis
Abstract:  Recombinant yeast pyruvate kinase has been purifiedfrom a strain of Saccharomyces cerevisiae expressing theenzyme to very high levels. Expression was from a multicopyplasmid under the control of the yeast phosphoglycerate kinasepromoter. The gene was expressed in the absence of thegenomically encoded pyruvate kinase, using a strain of yeastin which the pyruvate kinase gene has been disrupted by theinsertion of the yeast Ura3 gene. The purification procedureminimised proteolytic artefacts and enabled the covenientpurification of 15-20 mg enzyme from 1 l culture. The purifiedenzyme was characterised by a high specific activity and by alack of proteolytic degradation. Two active-site mutants ofyeast pyruvate kinase have been produced, expressed andcharacterised in this system and preliminary results aredescribed.
113                                 NAL Call. No.: 500 N21PRegulated expression of the GAL4 activator gene in yeastprovides 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 UnitedStates of America v. 88 (19): p. 8597-8601; 1991 Oct01. Includes references.
Language:  English
Descriptors: Saccharomyces cerevisiae; Strains; Yeasts; Geneexpression; Genetic regulation; Glucose; Inhibitors;Mutations; Promoters; Synergism; Transcription
Abstract:  Glucose (catabolite) repression is mediated bymultiple mechanisms that combine to regulate transcription ofthe GAL genes over at least a thousandfold range. We havedetermined that this is due predominantly to modest glucoserepression (4- to 7-fold) of expression of GAL4, the geneencoding the transcriptional activator of the GAL genes. GAL4regulation is affected by mutations in several genespreviously implicated in the glucose repression pathway; it isnot dependent on GALA or GAL80 protein function. GAL4 promotersequences that mediate glucose repression were found to liedownstream of positively acting elements that may be "TATAboxes." Two nearly identical sequences (10/12 base pairs) inthis region that may be binding sites for the MIG1 proteinwere identified as functional glucose-control elements. A 4-base-pair insertion in one of these sites causes constitutiveGAL4 synthesis and leads to substantial relief (50-fold) ofglucose repression of GAL1 expression. Furthermore, promoterdeletions that modestly reduce GAL4 expression, and thereforepresumably the amount of GAL4 protein synthesized, cause muchgreater reductions in GAL1 expression. These results suggestthat GAL4 works synergistically to activate GAL1 expression.Thus, glucose repression of GAL1 expression is due largely toa relatively small reduction of GAL4 protein levels caused byreduced GAL4 transcription. This illustrates how modestregulation of a weakly expressed regulatory gene can act as asensitive genetic switch to produce greatly amplifiedresponses to environmental changes.
114                                 NAL Call. No.: 381 B523Rotation and interactions of genetically expressed cytochromeP-450IA1 and NADPH-cytochrome P-450 reductase in yeastmicrosomes.Iwase, T.; Sakaki, T.; Yabusaki, Y.; Ohkawa, H.; Ohta, Y.;Kawato, S. Washington, D.C. : American Chemical Society; 1991Aug27. Biochemistry v. 30 (34): p. 8347-8351; 1991 Aug27. Includes references.
Language:  English
Descriptors: Saccharomyces cerevisiae; Microsomes; Cytochromep-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 yeastmicrosomes. The ratio of P-45OIA1 to the reductase was about17:1 and 1:2 without and with coexpression of the reductase,respectively. Rotational diffusion of P-450IA1 was examined byobserving the flash-induced absorption anisotropy, r(t), ofthe heme-CO complex. In only P-450Ia1-expressed microsomes,28% of P-450IA1 was rotating with a rotational relaxation timeof about 1200 microseconds. The mobile population wasincreased to 43% by the presence of the coexpressed reductase,while rotational relaxation time was not changedsignificantly. Increased concentration of KCl from 0 to 1000mM caused considerable mobilization of P-450IA1. The resultsdemonstrate a proper incorporation of P-450IA1 molecules intoyeast microsomal membranes. The significant mobilization ofP-450IA1 by the presence of reductase suggests a possibletransient association of P-450IA1 with the reductase.
115                                 NAL Call. No.: 381 J824Saccharomyces 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 andMolecular 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 acidsequences
Abstract:  The elongation factor 2 (EF-2) genes of the yeastSaccharomyces cerevisiae have been cloned and characterizedwith the ultimate goal of gaining a better understanding ofthe mechanism and control of protein synthesis. Two genes(EFT1 and EFT2) were isolated by screening a bacteriophagelambda yeast genomic DNA library with an oligonucleotide probecomplementary to the domain of EF-2 that contains diphthamide,the unique posttranslationally modified histidine that isspecifically ADP-ribosylated by diphtheria toxin. AlthoughEFT1 and EFT2 are located on separate chromosomes, the DNAsequences of the two genes differ at only four positions outof 2526 base pairs, and the predicted protein sequences areidentical. Genetic deletion of each gene revealed that atleast one functional copy of either EFT gene is required forcell viability. Messenger RNA levels of yeast EF-2 parallelcellular growth and peak in mid-log phase cultures. The EF-2protein sequence is strikingly conserved through evolution.Yeast EF-2 is 66% identical to, and shares over 85% homologywith, human EF-2. In addition, yeast and mammalian EF-2 shareidentical sequences at two critical functional sites: (i) thedomain containing the histidine residue that is modified todiphthamide and (ii) the threonine residue that isspecifically phosphorylated in vivo in mammalian cells bycalmodulin-dependent protein kinase III, also known as EF-2kinase. Furthermore, yeast EF-2 also contains the Glu-X-X-ArgX-Ile-Thr-Ile "effector" sequence motif that is conservedamong all known elongation factors, and its GTP-binding domainexhibits strong homology to the G-domain of Escherichia colielongation factor Tu (EF-TU) and other G-protein familymembers. Based upon these observations, we have modeled the Gdomain of the deduced EF-2 protein sequence to the solvedcrystallographic structure for EF-TU.
116                                NAL Call. No.: 442.8 Z34The Saccharomyces cerevisiae SPT14 gene is essential fornormal expression of the yeast transposon, Ty, as well as forexpression of the HIS4 gene and several genes in the matingpathway.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; Geneexpression; Transcription; Genetic regulation; Mating; Genemapping; Gene location; Chromosomes; Restriction mapping;Messenger  RNA; Mutations
Abstract:  To investigate the role of the trans-actingtranscription factor encoded by the essential SPT14(SPT=suppressor of Ty insertion mutations) gene, we havecloned, mapped and sequenced the gene. From the analysis ofthe effect of spt14 mutations on expression of various genes,we conclude that the SPT14 product has an important role inactivation of Ty transcription as well as in the regulation ofother genes including HIS4 and several of the a- and alphaspecific mating type genes. Similarities in the phenotypes ofspt14 and spt13 mutants (suppression of Ty insertion mutationsbut not delta insertion mutations), lead to the suggestionthat the SPT14 gene and the previously characterizedSPT13/GAL11 gene might encode transcriptional regulators withrelated functions. Our current findings show that in contrastto SPT13/GAL11, which appears negatively to regulate Tytranscription, SPT14 plays a role in the activation of Tytranscription. Thus, despite the similarities in thesuppression phenotype exhibited by spt13 and spt14 mutants,SPT13/GAL11 and SPT14 probably differ in their transcriptionalroles.
117                                 NAL Call. No.: 500 N21PSelective induction of gene expression and second-messengeraccumulation in Dictyostelium discoideum by the partialchemotactic 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 UnitedStates 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 moldDictyostelium discoideum, cAMP induces chemotaxis andexpression of different classes of genes by means ofinteraction with surface cAMP receptors. We describe a cAMPderivative, 8-p-chlorophenylthioadenosine 3',5'-cyclicmonophosphate (8-CPT-cAMP), which inhibits cAMP-inducedchemotaxis at low concentrations but induces chemotaxis atsupersaturating concentrations. This compound, moreover,selectively activates expression of aggregative genes but notof postaggregative genes. 8-CPT-cAMP induces normal cGMP andcAMP accumulation but in contrast to cAMP, which increasesinositol 1,4,5-trisphosphate levels, 8-CPT-cAMP decreasesinositol 1,4,5-trisphosphate levels. The derivative inducesreduced activation of guanine nucleotide regulatory proteins,which may cause its defective activation of inositol 1,4,5-trisphosphate production. Our data suggest that disruption ofinositolphospholipid signaling impairs chemotaxis andexpression of a subclass of cAMP regulated genes.
118                                 NAL Call. No.: QH426.C8Sequence analysis of the ARG7 gene of Schizosaccharomycespombe coding for argininosuccinate lyase. Expression of thegene 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.  Includesreferences.
Language:  English
Descriptors: Yeasts; Saccharomyces cerevisiae; Genetic code;Nucleotide sequences; Amino acid sequences; Argininosuccinatelyase; Enzyme activity; Gene expression; Gene mapping
119                                NAL Call. No.: 442.8 G28Site-specific recombination determined by I-SceI, amitochondrial group I intron-encoded endonuclease expressed inthe 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.  Includesreferences.
Language:  English
Descriptors: Saccharomyces cerevisiae; Introns; Mitochondrial DNA; Nucleases; Gene expression; Nuclei; Recombination; Dnamodification; Enzyme activity; Dna repair; Gene conversion;Deletions
Abstract:  The Saccharomyces cerevisiae mitochondrialendonuclease I-SceI creates a double-strand break as theinitiating step in the gene conversional transfer of theomega+ intron to omega- DNA. We have expressed a galactoseinducible synthetic I-SceI gene in the nucleus of yeast thatalso carries the I-SceI recognition site on a plasmidsubstrate. We find that the galactose-induced I-SceI proteincan be active in the nucleus and efficiently catalyzerecombination. With a target plasmid containing direct repeatsof the Escherichia coli lacZ gene, one copy of which isinterrupted by a 24-bp cutting site, galactose inductionproduces both deletions and gene conversions. Both thekinetics and the proportion of deletions and gene conversionsare very similar to analogous events initiated by a galactoseinducible HO endonuclease gene. We also find that, in a rad52mutant strain, the repair of double-strand breaks initiated byI-SceI and by HO are similarly affected: the formation ofdeletions is reduced, but not eliminated. Altogether, theseresults suggest either that the two endonucleases act in thesame way after double-strand break formation or that the twoendonucleases are not involved in subsequent steps.
120                                 NAL Call. No.: 450 P692Slow-growth phenotype of transgenic tomato expressingapoplastic invertase. Dickinson, C.D.; Altabella, T.;Chrispeels, M.J.Rockville, Md. : American Society of Plant Physiologists; 1991Feb. 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 mosaiccaulimovirus; Symptoms; Light; Photosynthates; Translocation
Abstract:  The growth of transgenic tomato (Lycopersiconesculentum) plants that express in their apoplast yeastinvertase under the control of the cauliflower mosaic virus35S promoter is severely inhibited. The higher the level ofinvertase, the greater the inhibition of growth. A secondphenotypic characteristic of these transgenic plants is thedevelopment of yellow and necrotic spots on the leaves, andleaf curling. Again the severity of the symptoms is correlatedwith the level of invertase. These symptoms do not develop inshaded leaves indicating the need for photosynthesis. Keepingthe plants in the dark for a prolonged period (24 hours)results in the disappearance of leaf starch from the controlplants, but not from the plants with apoplastic invertase.These results are consistent with the interpretation thatapoplastic invertase prevents photosynthate export from sourceleaves and that phloem loading includes an apoplastic step.
121                                 NAL Call. No.: QH426.C8Structure and express of a plastid-encoded groEL homologousheat-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.  Includesreferences.
Language:  English
Descriptors: Rhodophyta; Chloroplast genetics; Plastids; Dna;Genes; Plant proteins; Nucleotide sequences; Amino acidsequences; Transcription; Gene expression; Translation; Heatshock; Restriction mapping
Abstract:  A gene homologous to the E. coli groEL locus wasidentified on the plastid genome of the unicellular red algaCyanidium caldarium strain 14-1-1 (synonym: Galdieriasulphuraria). The complete nucleotide sequence was determinedand compared to bacterial- and nuclear-encoded counterparts ofhigher plants. At the amino-acid level the C. caldarium geneshows 70% homology to the corresponding gene of thecyanobacterium Synechococcus and 52% homology to nuclearencoded counterparts of higher plants, respectively. Northernand Western blot experiments were used to investigate thedependence of the transcript- and protein-level on culturetemperature and heat shock.
122                                  NAL Call. No.: 472 N21SWI6 protein is required for transcription of the periodicallyexpressed 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.  Includesreferences.
Language:  English
Descriptors: Yeasts; Dna; Gene expression; Proteins;Transcription
Abstract:  IN budding yeast many genes are expressed undercell-cycle control in late G1. These include a large group ofDNA synthesis genes, the HO gene involved in mating-typeswitching, C7S1 (chitinase) and also CLN1 and CLN2 (ref. 4)encoding G1 cyclins. Two factors, encoded by the SW14 and SW16genes, 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 notfound upstream of the DNA synthesis genes, which instead havea hexamer element, ACGCGT (MCB), an MluI restriction site,that is recognized by a cell-cycle regulated transcriptioncomplex DSC1 (ref. 1). This MluI-activation system consistingof the MCBs and DSC1 is conserved in fission yeast where aDSC1-like complex controls the cdc22+ ribonucleotide reductasegene. The Schizosacharomyces pombe cdc10+ gene encodes acomponent of DSC1 (ref. 10) and, significantly, this hashomology with both the Swi4 and Swi6 proteins. Here we showthat Swi6 is an essential component of DSC1 and that deletionof SW16 impairs the cell-cycle regulation of the DNA synthesisgenes, as well as CLN1 and CLN2. Thus Swi6 is the commonfactor in regulation of all the above genes and may thereforebe responsible for the timing of their expression in late G1.
123                               NAL Call. No.: QH442.A1G4Synthesis and expression of gene encoding tuna, pigeon, andhorse 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; Nucleotidesequences; Amino acid sequences
Abstract:  Genes encoding tuna, pigeon, and horse cytochromesc- were constructed with synthetic oligodeoxyribonucleotideshaving preferred codons and portions of the iso-1-cytochromec-encoding gene from the yeast Saccharomyces cerevisiae. Thegenes were ligated into an expression vector, which containsthe normal 5'- and 3'-untranslated regions of the yeast iso-1-cytochrome c gene, and were integrated in single copy into thechromosome. Yeast strains were also constructed with multipleintegrated copies of the pigeon gene. The heterologous andnormal mRNA levels of the single-copy strains were equivalent.Although the N-terminal methionines were completely cleaved inthe heterospecific proteins, the levels of trimethylation ofLys72 and acetylation of N-terminal glycines ranged from39-78% and 10-70%, respectively. Horse cytochrome c wasproduced at a nearly normal level, whereas the pigeon and tunacytochromes c were produced at approx. 40% of the normallevels. The levels of the cytochromes c and growth of themutant yeast strains indicated that the heterospecificcytochromes had approx. 50% specific activity in vivo.
124                                 NAL Call. No.: 500 N484Systems and approaches for expression and secretion ofheterologous proteins in the filamentous fungus Aspergillusniger 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; Geneexpression; Genetic engineering; Food industry; Industrialmicrobiology
125                                NAL Call. No.: SB599.P45Temperature and genotype interactions in the expression ofhost 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; Hostspecificity; Disease resistance; Genotypes; Gene expression;Virulence; Genes; Environmental factors; Air temperature;Genotype environment interaction
Abstract:  The influence of temperature and genotype oninfection types determined by 13 gene-for-gene interactionsbetween lettuce and the downy mildew fungus, Bremia lactucae,was evaluated. Lettuce seedlings containing each of 13resistance (Dm) genes were inoculated with isolates of B.lactucae expressing various combinations of the matchingavirulence genes, and incubated at 5, 10, 15, or 22 degrees C.Resistance, assessed by the absence of sporulation, becameless effective at lower temperatures for Dm6, Dm7, Dm11, Dm15,and Dm16. No change was observed for resistance conditioned byeight other gene-for-gene interactions. Histologicalexamination indicated that the extent of fungal development inhost tissue was dependent on the interacting genotypes and ontemperature. Manipulation of interactions involving Dm7 andDm11, using temperature-shift incubation regimes, indicatedthat continued and cumulative exposure to permissivetemperatures was required for effective resistance andsuggested that the determinants of specificity are present inmost host cells and are expressed throughout fungaldevelopment.
126                               NAL Call. No.: 448.3 J823The thn mutation of Schizophyllum commune, which suppressesformation aerial hyphae, affects expression of the Sc3hydrophobin gene.Wessels, J.G.H.; Vries, O.M.H. de; Asgeirsdottir, S.A.;Springer, J. Reading : Society for General Microbiology; 1991Oct.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 thebasidiomycete Schizophyllum commune suppresses the formationof aerial hyphae in the monokaryon and, if present as a doubledose, the formation of both aerial hyphae and fruit-bodies inthe dikaryon. In the monokaryon, the mutation preventsaccumulation of mRNA of the Sc3 gene, and in the dikaryon italso prevents the accumulation of fruiting-specific mRNAs,including mRNAs of the Sc1 and Sc4 genes, which are homologousto the Sc3 gene. These three genes code for hydrophobins, afamily of small hydrophobic cysteine-rich proteins. In the thnmonokaryon, the only detectable change in synthesized proteinsis the disappearance of an abundant protein of apparent Mr =28 K from the culture medium and from the cell walls. Proteinsequencing shows that this is the product of the Sc3 gene. TheSc3 hydrophobin is present in the walls of aerial hyphae as ahot-SDS-insoluble complex. Submerged hyphae excrete largeamounts of the hydrophobin into the medium.
127                               NAL Call. No.: QD341.A2N8Transcript mapping reveals different expression strategies forthe 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; Geneticregulation; Gene expression; Rna; Polymerase chain reaction;Molecular mapping; Nucleotide sequences
Abstract:  We have characterised the major transcripts of theCzech isolate of wheat dwarf virus (WDV-CJI) which show thatWDV uses two different mechanisms for expressing overlappingopen reading frames (ORFs). Mapping of the virion sense RNAsidentified a single polyadenylated transcript of 1.1kbspanning the overlapping ORFs V1 and V2 which encode cell-cellspread functions and the coat protein respectively. Thisfinding distinguishes WDV from other monocot-infectinggeminiviruses studied so far which were shown to encode two 3'co-terminal transcripts capable of expressing either the V1 orV2 ORF. A survey of codon usage at the junction between the V1and V2 ORF has led us to propose that translational frameshifting analogous to that in the yeast Ty element may occur.Analysis of polymerase chain reaction (PCR) amplifiedcomplementary sense cDNA clones has revealed the presence ofmature spliced and unspliced RNAs which could encode productsof an intron mediated C1:C2 ORF fusion or the C1 ORF productalone. Mapping of the 51 and 31 extremities of the major WDVencoded transcripts has allowed us to identify putativetranscription regulatory sequences and the presence ofmultiple overlapping transcripts may suggest temporalregulation of transcription.
128                                 NAL Call. No.: QH426.C8Transient expression of firefly luciferase in protoplasts ofthe 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.  Includesreferences.
Language:  English
Descriptors: Chlorella ellipsoidea; Protoplasts; Genetictransformation; Luciferase; Reporter genes; Gene expression;Gene mapping
129                                 NAL Call. No.: QH426.C8Transient expression of genes in the oomycete Phytophthorainfestans 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.  Includesreferences.
Language:  English
Descriptors: Bremia lactucae; Phytophthora infestans; Genetictransformation; Gene expression; Beta-glucuronidase; Reportergenes; Vectors; Plant pathogenic fungi
Abstract:  Vectors containing fusions between the reportergene beta-glucuronidase (GUS) and transcriptional regulatorysignals from the ham34 and hsp70 genes of the oomycetepathogen, Bremia lactucae, were introduced into protoplasts ofthe related fungus Phytophthora infestans. Transientexpression of the GUS gene was detected when DNA wasintroduced into protoplasts of P. infestans by treatments withpolyethylene glycol and CaCl(2), with cationic liposomes, orby electroporation. After optimization of each procedure,using the transient expression assays, cationic liposomes wereidentified as the superior method for DNA uptake. Vectorscontaining the 5'hsp70 sequences and 3'ham34 sequencesresulted in the maximum level of GUS activity. Theidentification of functional vectors for transformation, andof optimal methods for introducing DNA, should assist inachieving stable transformation of P. infestans and otheroomycete the fungi.
130                                NAL Call. No.: 464.8 P56Transient expression of the beta-glucuronidase gene introducedinto Uromyces appendiculatus uredospores by particlebombardment.Bhairi, S.M.; Staples, R.C.St. Paul, Minn. : American Phytopathological Society; 1992Sep. Phytopathology v. 82 (9): p. 986-989; 1992 Sep.  Includesreferences.
Language:  English
Descriptors: Phaseolus vulgaris; Uromyces appendiculatus;Infectivity; Genetic regulation; Gene expression; Genetictransformation; Beta-glucuronidase
Abstract:  Plasmids carrying the beta-glucuronidase gene (GUS)under the control of the promoter from the previously clonedgene specific for infection structure, INF24, were constructedby inserting the INF24 promoter in front of the GUS genecarried in the Bluescript vector. These plasmid DNAs were thenintroduced into uredospores of the bean rust fungus, Uromycesappendiculatus; the biolistic particle delivery system wasused. With germination and differentiation on collodionmembranes, GUS activity assayed histochemically with X-glucwas evident only in those germlings that were bombarded withthe plasmid containing the entire promoter region; it was notevident in those that were mock-bombarded or bombarded withthe plasmid lacking the promoter.
131                               NAL Call. No.: QH442.A1G4 Atwin-reporter vector for simultaneous analysis of expressionsignals of divergently transcribed, contiguous genes infilamentous 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. vanAmsterdam : 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; Geneexpression; Nitrate reductase; Nitrite reductase
Abstract:  To analyze the promoter region(s) of divergently.Transcribed fungal genes, a twin-reporter vector wasconstructed. This vector contains two divergently orientedreported genes, encoding Escherichia coli beta-glucuronidase(uidA) and E. coli beta-galactosidase (lacZ). Terminatorregions of the Aspergillus nidulans nitrate and nitritereductase-encoding genes, niaD and niiA, respectively, havebeen cloned 3' to the reporter genes to ensure proficienttranscription termination of the reporter genes. The reportergenes have been separated by a unique NotI restriction site,which can be used for the insertion of expression signals. Amutant argB selection marker has been introduced in order toobtain A. nidulans transformants with a single copy of thevector integrated at the argB locus. The use of the vector wasdemonstrated by insertion of the A. nidulans niaD-niiAintergenic region and analysis of A. nidulans transformantsobtained with this construct. Control of expression of bothreporter genes was found to be in accordance with previouslypublished data on control of nitrate assimilation [Cove, Biol.Rev. 54 (1979) 291-327].
132                                 NAL Call. No.: 381 J824Two yeast genes encoding calmodulin-dependent protein kinases.Isolation, sequencing, and bacterial expressions of CMK1 andCMK2.Ohya, Y.; Kawasaki, H.; Suzuki, K.; Londesborough, J.; Anraku,Y. Baltimore, Md. : American Society for Biochemistry andMolecular 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 Saccharomycescerevisiae that both encode a calmodulin-dependent proteinkinase (CaM kinase). The CMK1 gene has been cloned byhybridization using an oligonucleotide probe synthesized onthe 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 beenisolated by screening at low stringency with a CMK1 fragmentas a probe. The CMK2 product expressed in bacteria shows Ca2+-and CaM-dependent protein kinase activity, indicating thatCMK2 also encodes a CaM kinase. The CMK1 and CMK2 productsexpressed in bacteria were found to have different biochemicalproperties in terms of autoregulatory activity and preferencefor yeast CaM or bovine CaM for maximal activity. Antibodyraised against a peptide fragment of the CMK1 protein crossreacts with the CMK2 product. Immunoblotting with thisantibody indicated that the CMK1 and CMK2 products haveapparent molecular masses of 56 and 50 kDa, respectively, inyeast cells. The predicted amino acid sequences of the two CMKproducts exhibit highest similarity with mammalian calmodulindependent multifunctional protein kinase II (CaM kinase II):the similarity within the N-terminal catalytic domain is about40%, whereas that within the rest of the sequence is 25%.These data indicate that yeast has two kinds of genes encodingCaM kinase isozymes whose structural and functional propertiesare closely related to those of mammalian caM kinase II.Another gene may be substituted for function of the CMK1 andCMK2 kinase in vivo, since elimination of both kinase genes isnot lethal.
133                                 NAL Call. No.: QH426.C8Ubiquitin 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.  Includesreferences.
Language:  English
Descriptors: Saccharomyces cerevisiae; Gene expression; Plantproteins; Genes; Transcription; Messenger  RNA; Geneticregulation; Heat shock; Methyl methanesulfonate; Mutagens;Starvation; Refeeding
Abstract:  It has previously been shown that the yeastubiquitin genes UBI1, 2 and 3 are strongly expressed duringthe log-phase of batch culture growth, whereas the U8I4 geneis weakly expressed. We found that heat shock, treatment withDNA-damaging agents, starvation, and the feeding of starvedcells all transiently induced UBI4. These results suggest thatUBI4 is induced whenever a change in culture conditionsdictates a dramatic shift in cellular metabolism, and thatUBI4 expression returns to lower levels once cellularmetabolism has adapted to the new conditions. In contrast, allof the treatments tested, except starvation, transientlyrepressed the UBI1, 2 and 3 genes. Although starvation alsorepressed UBI1, 2 and 3 its effect was not transient, andexpression only recovered upon the addition of fresh media.These results, together with others presented here, suggestthat high levels of UBI1, 2 and 3 expression are dependantupon ongoing cell growth, and that treatments which slow orstop growth repress their expression.
134                               NAL Call. No.: QH442.A1G4Vectors for the expression and analysis of DNA-bindingproteins 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 yeastcentromere plasmids with the LEU2 gene for selection in yeast,and pUC19 sequences for growth in Escherichia coli. Allcontain the GAL1 promoter directing transcription into amultiple 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. Spacingbetween the ATG and the MCS is variable, to facilitate thecloning of gene fragments in the appropriate reading frame.Nine of the plasmids also contain the strong transcriptionalactivator from the herpes simplex virus VP16 gene, joineddownstream from the MCS. In these nine vectors, all possiblecombinations of reading frames are available. The suitabilityof these plasmids for the expression and analysis of DNAbinding domains is tested by cloning into them fragments ofthe yeast HSF1 gene, encoding the heat shock transcriptionfactor (HSF). The regulation of reporter gene expression bythe chimeric HSF-VP16 fusions is described, as is the utilityof these vectors for other applications.
135                                NAL Call. No.: QH506.E46Virus-like genetic organization and expression strategy for adouble-stranded RNA genetic element associated with biologicalcontrol 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; Biologicalcontrol; Hypovirulence; Rna; Nucleotide sequences
Abstract:  The complete nucleotide sequence of the largestdouble-stranded (ds) RNA present in hypovirulent strain EP713of the chestnut blight pathogen, Cryphonectria parasitica, wasdetermined and the predicted genetic organization wasconfirmed by translational mapping analysis. The deduced RNAsequence was 12 712 bp in length, excluding the terminalpoly(A):poly(U) homopolymer domain. The strand terminatingwith 3'-poly(A) contained two contiguous large open readingframes (ORF A and ORF B) beginning at nucleotide residues496-498 and extending to nucleotide positions 11 859-11 861.The junction between ORF A and ORF B consisted of the sequence5'-UAAUG-3', where UAA served as the termination codon for ORFA 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 anautocatalytic event mediated by p29 (Choi et al., 1991). Asimilar autocatalytic event was observed during in vitrotranslation of ORF B (3165 codons in length) resulting in therelease of a 48 kd polypeptide from the amino-terminal portionof the ORF B-encoded polyprotein. These results are discussedin terms of the opportunities they provide for elucidating themolecular basis of transmissible hypovirulence and possibleorigins of hypovirulence-associated dsRNAs.
136                                 NAL Call. No.: 381 J824VMA11, a novel gene that encodes a putative proteolipid, isindispensable for expression of yeast vacuolar membrane H+-ATPase activity. Umemoto, N.; Ohya, Y.; Anraku, Y.Baltimore, Md. : American Society for Biochemistry andMolecular 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; Nucleotidesequences
Abstract:  A gene, VMA11, is indispensable for expression ofthe vacuolar membrane H+-ATPase activity in the yeastSaccharomyces 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 genomicDNA library by complementation of the vma11 mutation. Thenucleotide sequence of the gene predicts a hydrophobicproteolipid of 164 amino acids with a calculated molecularmass of 17,037 daltons. The deduced amino acid sequence shows56.7% identity, and significant coincidence in amino acidcomposition with the 16-kDa subunit c (a VMA3 gene product) ofthe yeast vacuolar membrane H+-ATPase. VMA11 and VMA3 on amulticopy plasmid did not suppress the vma3 and vma11mutation, respectively, suggesting functional independence ofthe two gene products. Biochemical detection of the VMA11 geneproduct was unsuccessful, but vacuoles in the VMA11-disruptedcells were not assembled with either subunit c or subunits aand b of the H+-atpase, resulting in defects of the activityand in vivo vacuolar acidification.
137                               NAL Call. No.: 442.8 J828Yeast cell cycle protein CDC48p shows full-length homology tothe mammalian protein VCP and is a member of a protein familyinvolved in secretion, peroxisome formation, and geneexpression.Frohlich, K.U.; Fries, H.W.; Rudiger, M.; Erdmann, R.;Botstein, D.; Mecke, D. New York, N.Y. : RockefellerUniversity 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 acidsequences; Restriction mapping; Cell division;Immunocytochemistry; Comparisons; Animal proteins; Mammals;Microsomes
Abstract:  Yeast mutants of cell cycle gene cdc48-1 arrest aslarge budded cells with microtubules spreading aberrantlythroughout the cytoplasm from a single spindle plaque. Thegene was cloned and disruption proved it to be essential. TheCDC48 sequence encodes a protein of 92 kD that has an internalduplication of 200 amino acids and includes a nucleotidebinding consensus sequence. Vertebrate VCP has a 70% identityover the entire length of the protein. Yeast Sec18p andmammalian N-ethylmaleimide-sensitive fusion protein, which areinvolved in intracellular transport, yeast Pas1p, which isessential for peroxisome assembly, and mammalian TBP-1, whichinfluences HIV gene expression, are 40% identical in theduplicated region. Antibodies against CDC48 recognize a yeastprotein of apparently 115 kD and a mammalian protein of 100kD. Both proteins are bound loosely to components of themicrosomal fraction as described for Sec18p and Nethylmaleimide-sensitive fusion protein. This similaritysuggests that CDC48p participates in a cell cycle functionrelated to that of N-ethylmaleimide-sensitive fusionprotein/sec18p in Golgi transport.                         AUTHOR INDEX
Achstetter, T.  44Alberghina, L.  107Allen, J.  21Allen, S.C.  112Altabella, T.  120Amore, R.  14Anderson, J.A.  67Anraku, Y.  72, 96, 132, 136Archer, D.B.  111Arimura, H.  15Asgeirsdottir, S.A.  126Atomi, H.  77Audino, D.C.  7Auer, B.  83Aust, S.D.  74Baetselier, A. de  38Bandlow, W.  64Barbetti, M.J.  37Barnes, C.A.  56Bartley, G.E.  93Begdadi-Rais, C.  32Beier, D.  63Beier, H.  63, 106Berberian, G.E.  69Berka, R.M.  124Berka, T.R.  21Bernard, M.  32Bertheau, Y.  17Bhairi, S.  10Bhairi, S.M.  130Birchmeier, C.  62Blachly-Dyson, E.  108Black-Schaefer, L.  89Blaiseau, P.L.  17Blomqvist, K.  12Bodley, J.W.  115Bominaar, A.A.  117Bonneaud, N.  16Bonner, J.J.  134Borgford, T.J.  42Botstein, D.  137Bourbonnais, Y.  75Bouzayen, M.  79Brandt, R.  117Breeden, L.  122Bron, C.  32Brown, L.M.  128Bruenn, J.A.  66Brygoo, Y.  17Buloichik, A.A.  99Bunkers, G.J.  60Bussink, H.J.D.  46Buxton, F.P.  46Cannons, A.C.  49Cano-Canchola, C.  31Caput, D.  13Carmo-Fonseca, M.  41Carr, C.  6Carron, C.P.  6Cauwelaert, F. van  38Ceccarelli, A.  117Chamovitz, D.  93Chiang, C.C.  54, 70Choi, G.H.  29, 91, 135Chotai, D.  4Chrispeels, M.J.  120Ciriacy, M.  14Citavicius, D.  84Clements, J.M.  123Colicelli, J.  62Collins, W.J.  37Colombini, M.  108Conneely, O.M.  98Connerton, I.F.  23Corradin, G.  32Cox, B.  127Cregg, J.M.  76Curtis, P.J.  40Danoff, A.  75Davies, R.W.  55Davio, M.  18De Zoysa, P.A.  23Decroupette, I.  118Deforce, L.  82Dekker, E.L.  127Delpech, B.  13Denis, C.L.  7Deschenes, R.J.  19Deutekom, J.C.T. van  131Dickinson, C.D.  120Dignard, D.  48Dimarcq, J.L.  44Dittrich, H.  94Dixon, J.E.  19Dubois, E.  20Dujon, B.  119Dumont, X.  13Durand, R.  95Duronio, R.J.  6Dykes, C.W.  4Eichinger, L.  35Engelke, D.R.  58Erdmann, R.  137Evans, C.F.  58Farrens, D.  82Fasel, N.  32Fassler, J.S.  116Feaver, W.J.  11Ferguson, C.  69Ferrara, P.  13Fevre, M.  95Fingar, S.A.  123Firtel, R.A.  80Fleig, U.N.  50Fonzi, W.  31Forlani, N.  107Forte, M.  108Fothergill-Gilmore, L.A.  112Fraser, J.  133French, F.S.  4Fried, M.  101Friedrich, B.  53Fries, H.W.  137Frohlich, K.U.  137Fujimura, T.  65Furuya, M.  82Gaber, R.F.  67Gaisne, M.  30Gaymard, F.  16Germain, D.  48Giallongo, A.  101Gillespie, D.J.  37Gingerich, G.  116Gleason, W.B.  115Gold, M.H.  85Gomi, K.  26Goodhue, C.T.  123Gordon, J.I.  6Gough, K.H.  57Gralla, E.B.  1Gray, W.  116Greaves, P.A.  131Greenberg, M.L.  105Grierson, D.  79Griggs, D.W.  113Grignon, C.  16Grison, R.  17Grivell, L.A.  78Guan, K.  19Guillemot, J.C.  13Haas, M.J.  21Haastert, P.J.M. van  117Haber, J.E.  119Hadwiger, J.A.  80Hadwiger, L.A.  54, 70Haguenauer-Tsapis, R.  2Hahlbrock, K.  104Haid, A.  64Halpert, J.R.  68Hambidge, S.J.  34Hamilton, A.J.  79Hanau, R.M.  71Hara, S.  26Harkki, A.  92Hebraud, M.  95Heinen, Ute  90Hermann, H.  64Hickey, D.R.  123Hinnen, A.  2Hirschberg, J.  93Hobden, A.N.  4Hoffmann, J.A.  44Hohmann, S.  103Hohn, T.M.  51Hollenberg, C.P.  14Hondel, C.A.M.J.J. van  131Horaitis, O.  81Hori, K.  22Horiuchi, H.  59Horovitz, D.  54Hosaka, K.  36, 47Hosokawa, Y.  123Huffel, C. van  20Huprikar, S.S.  67Hurt, E.C.  41Icho, T.  65Iida, H.  96Iida, N.  49Irie, M.  59Ishida, Y.  15Ito, N.  82Iwase, T.  114Jagadish, M.N.  57Jansen, R.P.  41Jarvis, E.E.  128Jayaraman, K.  123Jeenes, D.J.  111Johnson, A.L.  28, 122Johnson, T.M.  73Johnston, G.C.  56Johnston, J.R.  23Johnston, L.H.  28, 122Johnston, M.  113Joniau, M.  38Joos, H.J.  104Joris, B.  118Judelson, H.S.  125, 129Kaiser, P.  83Kajiwara, S.  22Kalmar, G.B.  42Kanamori, J.  109Karkhoff-Schweizer, R.R.  105Katayose, Y.  22Kato, R.  9Kawabe, H.  15Kawasaki, H.  132Kawato, S.  114Kedzie, K.M.  68Kelly, B.L.  105Keppi, E.  44Kern, H.  41Kim, C.S.  109Kinal, H.  66Kinghorn, J.R.  131Kirk, N.  87Kirkegaard, K.  34Kitamoto, K.  26Kito, M.  109Klar, A.J.S.  25Knowles, J.  12, 92Kochian, L.V.  67Kodaki, T.  36Komel, R.  3Kondo, J.  77Korte, A.  5, 102Kotter, P.  14Kotula, L.  40Krabusch, M.  8Kues, U.  53Kunz, C.  17Kuster, C.  14Kutchan, T.M.  94Kuyvenhoven, A.  131Labbe-Bois, R.  30Lacroute, F.  16Langner, C.A.  6Lapeyre, B.  41LaPorte, D.C.  115Larson, T.G.  91Lecocoq, J.P.  44Lee, J.P.  116Lee, J.Y.  58Legoux, R.  13Legrain, M.  44Lehtonen, H.  41Li, J.K.K.  73Linz, J.E.  33Livingston, D.M.  115Lockridge, O.  18Loison, G.  13Londesborough, J.  132London, A.  45Loper, J.C.  97Loppes, R.  118Lottspeich, F.  102Lowndes, N.F.  28, 122Lubahn, D.B.  4Lucas, W.J.  67Lukins, H.B.  110Luo, S.  24Luu, H.A.  133MacKenzie, D.A.  111Macreadie, I.G.  81Madgwick, P.J.  52Maid, U.  121Martegani, E.  107Masaki, A.  15Massey, V.  18McAlister-Henn, L.  43McCourt, J.D.  89McDonnell, D.P.  98McKee, E.E.  89McKown, R.L.  39McNally, T.  112Mecke, D.  137Meskauskas, A.  84Messenguy, F.  20Michaelis, U.  5, 102Michelmore, R.W.  125, 129Michels, R.  118Minard, K.I.  43Minet, M.  16Minnerly, J.C.  6Miyazawa, H.  22Monod, M.  2Motai, H.  15Muirhead, H.  112Mullineaux, P.M.  127Murakami, K.  15Murakami, S.  15Murcott, T.H.L.  112Nakajima-Shimada, J.  96Nakamoto, R.K.  61Nakano, E.  15Neculcea, J.  133Nevalainen, K.M.H.  92Nicolette, C.  62Nies, D.H.  53Nikawa, J.  36, 47Noegel, A.A.  35Nurse, P.  50Nuss, D.L.  29, 91, 135O'Malley, B.W.  98Oda, K.  77Oechsner, U.  64Ohgi, K.  59Ohkawa, H.  114Ohlrogge, J.B.  51Ohta, Y.  114Ohya, Y.  72, 132, 136Orlowski, M.  33Palmgren, M.G.  69Panaccione, D.G.  71Pauley, A.M.  6Payne, M.J.  110Pearlman, R.E.  11Pease, E.A.  74Pecker, I.  93Peery, R.B.  24Pel, H.J.  78Peng, S.  108Penttila, M.  12Penttila, M.E.  92Perentesis, J.P.  115Perie, F.H.  85Perrin, A.  119Peters, D.J.M.  117Petit, I.  44Phan, L.D.  115Philpot, R.M.  68Pines, O.  45Piper, P.W.  87Plessis, A.  119Poletti, A.  98Pouwels, P.H.  131Poyton, R.O.  89Pratt, K.A.  52Punt, P.J.  131Purvis, I.J.  4Qiu, H.  19Racher, K.I.  42Ramond, P.  13Rao, R.  61Reed, S.I.  27Reichhart, J.M.  44Rensing, C.  53Reymond, C.D.  32Reymond, P.  95Riggs, M.  62Roberts, I.N.  111Rodel, G.  5, 102Rodgers, L.  62Rogers, P.V.  24Rozman, D.  3Rudiger, M.  137Ruiz-Herrera, J.  31Russell, P.J.  34Rutherford, C.L.  24Ruusala, T.  86Saito, K.  18Saito, T.  22Sakaki, T.  114Salmon, J.M.  16Sato, T.  109Savin, K.W.  81Schaap, P.  117Schleicher, M.  35Schrader, W.T.  98Schweiger, M.  83Schweizer, E.  110Scolnik, P.A.  93Selmin, O.  24Sentenac, H.  16Serrano, R.  69Shah, J.  123Shapira, R.  29, 135Sherman, F.  123Shewry, P.R.  52Shields, D.  75Shire, D.  13Shishido, K.  22Shukla, D.D.  57Sierkstra, L.N.  100Silar, P.  1Silve, S.  2Singer, R.A.  56Slayman, C.W.  61Snaar-Jagalska, B.E.  117Solomonson, L.P.  49Song, P.S.  82Sosa, L.  31Springer, J.  126Stahl, U.  53Staples, R.C.  10, 130Steffan, J.S.  43Steinmuller, R.  121Storms, R.K.  133Strathmann, M.  80Sucic, J.F.  24Sudberry, P.E.  88Suihko, M.L.  12Sun, G.H.  72Suzuki, K.  132Sypherd, P.  31Szweykowska-Kulinska, Z.  106Tada, S.  26Takagi, M.  59Takahashi, K.  26Tamura, G.  26Tanaka, A.  77Tao, J.  66Tatsumi, H.  15Teeri, T.T.  92Teranishi, Y.  77Tessier, D.  48Thiele, D.J.  1, 18Thomas, D.Y.  48, 75, 133Thon, G.  25Tien, M.  74Togni, G.  2Tollervey, D.  41Tomizawa, K.I.  82Tschopp, J.F.  76Tsuji, F.I.  96Tsuji, R.F.  15Tsukagoshi, Y.  47Tsunasawa, S.  123Tulloch, P.A.  57Turi, T.G.  97Tzagoloff, A.  78Ueda, M.  77Umemoto, N.  136Utsumi, S.  109Valentine, J.S.  1Veale, R.A.  88Verbakel, J.M.A.  100Verdiere, J.  30Verkuylen, A.J.  81Verrips, C.T.  100Viaene, A.  38Viitanen, P.V.  93Villalba, J.M.  69Visser, J.  46Voluevich, E.A.  99Vries, O.M.H. de  126Vvolckaert, G.  38Ward, C.W.  57Warren, G.J.  39Watanabe, H.  59Watson, D.C.  23Weigel, N.L.  98Wessels, J.G.H.  126Whiteway, M.  48Whittaker, L.A.  57Wickner, R.B.  65Widner, W.R.  65Wiegand, R.C.  6Wigler, M.  62Wilkie, T.M.  80Williams, J.G.  117Wilson, E.M.  4Wittenberg, C.  27Wolf, K.  8Woolston, C.J.  127Xue, Y.  127Xuei, X.  10Yabusaki, Y.  114Yamashita, S.  36, 47Yamazoe, Y.  9Yang, Z.F.  71Yano, K.  59Yasumori, T.  9Yin, Y.  24Yoder, O.C.  10Yon, J.  101Yu, G.  116Zetsche, K.  121Zimmer, M.  8Zollner, A.  64                         SUBJECT INDEX
Acetylglutamic acid  20Acid phosphatase  2Actin  35, 91Adenosinetriphosphatase  61, 69, 136Adp  83Aegilops ovata  99Aegilops squarrosa  99Air temperature  125Alcohol dehydrogenase  7Alcohol oxidoreductases  88Alkanes  77Alleles  50, 103Alpha-amylase  26Alpha-galactosidase  100Alpha-lactalbumin  38Amino acid metabolism  107Amino 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, 137Anabolism  20Anaerobes  95Anaerobic conditions  30Androgens  4Androstenedione  3Animal proteins  137Antibacterial properties  44Antigens  32Arabidopsis thaliana  16, 63, 67, 69, 106Arginine  20Argininosuccinate lyase  118Asexual reproduction  71Aspergillus  46Aspergillus flavus  13Aspergillus nidulans  3, 55, 131Aspergillus niger  46, 55, 111, 124Aspergillus oryzae  15, 26Atp  110Autographa californica  74Bacterial proteins  39Beers  12Beta-fructofuranosidase  100, 120Beta-galactosidase  131Beta-glucuronidase  26, 60, 129, 130, 131Binding proteins  80, 117, 134Binding site  1, 15, 35, 49, 64, 83Biological control  135Biological development  22Biosynthesis  36, 51, 79, 82, 92, 94, 97, 107Blight  135Bremia lactucae  125, 129Brewing industry  12C-amp  7, 22, 24, 117Calcium ions  96Calmodulin  72, 132Candida albicans  6Candida tropicalis  77Carbohydrate metabolism  26Carboxy-lyases  12Carotenoids  93Castanea  91Castanea sativa  135Cauliflower mosaic caulimovirus  120Cell differentiation  10, 24Cell division  27, 50, 72, 83, 137Cell growth  96Cell ultrastructure  41Cell wall components  2Cellulolytic microorganisms  92Cellulose  92Cellulose digestion  92Characterization  38, 74, 112Chemical reactions  83Chitinase  17Chlorella ellipsoidea  128Chlorella vulgaris  49Chloroplast genetics  121Choline  36, 105Chromatin  11Chromosomes  116Chymosin  124Clones  20, 45, 49, 104Cloning  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, 137Coat proteins  57Cochliobolus lunatus  3Coenzyme a  6Cold resistance  39Colletotrichum graminicola  71Comparisons  91, 101, 137Conidia  71Controlling elements  25, 30, 64, 98Copper  1Crop damage  37Crop growth stage  37, 99Cryoprotectants  39Cryphonectria parasitica  29, 91, 135Cultivars  37, 54Cyamopsis tetragonoloba  100Cysteine  70Cytochrome b  5Cytochrome c  64, 123Cytochrome  9, 30, 97, 114Cytochrome-c oxidase  78Cytochromes  68Cytoplasmic inclusions  61Cytoplasmic inheritance  8, 99Deficiency  30Deletions  119Deuteromycotina  17Developmental stages  22, 31Diacetyl  12Dictyostelium  24, 32, 80, 117Diphosphatidylglycerols  105Disease resistance  37, 51, 54, 70, 125Dna  28, 31, 59, 62, 66, 70, 83, 93, 104, 121, 122Dna binding proteins  7, 30, 64, 116Dna hybridization  95Dna libraries  95Dna modification  119Dna repair  119Dna sequencing  11Dominance  54Dominant lethals  50Drug resistance  66Endomycetales  8, 25, 50, 86, 100Endoplasmic reticulum  2Enterobacter aerogenes  12Environmental factors  125Enzyme 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, 123Enzyme precursors  15Enzymes  97Ergosterol  97Escherichia coli  3, 21, 26, 43, 47, 57, 60Eschscholzia californica  94Ethylene  79Extracts  63Fermentation  76, 87Food industry  124Food research  109Fowls  98Freezing  39Fruiting  126Fungal antagonists  17Fungal diseases  29, 94, 99Fungal morphology  126Fungal protein  124Fungi  35, 55Fusarium oxysporum  17Fusarium oxysporum f.sp. pisi  54Fusarium solani f.sp. phaseoli  70Fusarium solani f.sp. pisi  70Fusarium sporotrichioides  51Gene conversion  119Gene 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, 136Gene location  25, 116Gene mapping  25, 80, 93, 116, 118, 128Gene transfer  21, 55, 83, 88, 100Genes  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, 133Genetic analysis  60, 70, 80, 91Genetic change  86Genetic code  20, 35, 40, 41, 46, 47, 58, 62, 77, 80, 84, 92,103, 109, 118Genetic control  29Genetic engineering  26, 40, 73, 98, 103, 109, 124Genetic markers  60Genetic regulation  7, 11, 17, 25, 27, 30, 36, 64, 78, 92, 97,103, 105, 106, 107, 110, 113, 116, 117, 127, 130, 133Genetic resistance  54, 99Genetic 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, 130Genomes  8, 104Genotype environment interaction  125Genotypes  125Gliadin  52Glucose  64, 113Glutamate dehydrogenase  23Glyceraldehyde-3-phosphate dehydrogenase  91Glycine max  93, 109Glycine soja  93Glycogen phosphorylase  24Growth  120Guanine  80Hansenula  88Heat shock  56, 62, 87, 121, 133, 134Hela cells  63Heme  30, 49, 64Hordeum vulgare  60Hormone receptors  98Horses  123Host parasite relationships  54, 70Host specificity  125Hydrolases  95Hygromycin b  66Hyperparasitism  17Hyphae  10, 126Hypovirulence  29, 91, 135Immunocytochemistry  109, 137Immunoglobulin structural genes  40In vitro  63Induced mutations  25, 48, 50, 78Induction  87, 95Industrial microbiology  124Infections  10Infectivity  130Inhibition  120Inhibitors  113Injuries  104Inositol phosphates  117Introns  10, 22, 91, 106, 119Ion transport  67Ions  53Isocitrate lyase  77Isoenzymes  9, 73, 74Isolation  6, 132Isoleucine  42Kabatiella caulivora  37Kinases  20Kinetics  33, 42, 105Klebsiella  12Laccase  85, 91Lactuca sativa  125Lanosterol  97Lentinula edodes  22Ligases  42, 51, 105, 110Light  120Lignin  73, 85Ligninolytic microorganisms  73Lines  54Literature reviews  27, 55, 88, 92Loci  25, 86Luciferase  128Luminescence  96Lycopersicon esculentum  79, 120Lysozyme  111Malate dehydrogenase  43Mammals  68, 75, 137Man  4, 9, 83Manganese  74, 85Marker genes  66Mating  25, 86, 116Meiosis  25Membranes  105, 108, 136Mercury  53Messenger  RNA  5, 10, 17, 25, 26, 33, 34, 63, 70, 78, 91,101, 104, 116, 123, 133Metabolism  12Metal ions  1Methanol  76, 87, 88Methionine  107Methyl methanesulfonate  133Methylation  31Mice  40, 101Microbial degradation  85Microbial proteins  77Microsomes  114, 137Mildews  99Mineralization  85Mitochondria  5, 20, 43, 89, 102, 105, 110Mitochondrial  DNA  8, 78, 119Mitochondrial genetics  8, 78Molds  117Molecular biology  92Molecular conformation  63, 108Molecular genetics  79, 94Molecular mapping  127Molecular weight  70Mucor  31, 33Multigene families  137Multiple genes  63Mutagenesis  112Mutagens  133Mutants  11, 20, 25, 35, 48, 50, 67, 78, 82, 92, 103, 108,110, 126 Mutations  8, 36, 63, 108, 113, 116, 126Mutator genes  8Mycelium  22Mycotoxins  48, 66, 84Myo-inositol  36, 105Myristic acid  6Nad  14Nadh  14Nadp  23Nadph  18Nadph-cytochrome-c2 reductase  114Neocallimastix  95Neurospora crassa  91Nicotiana rustica  106Nicotiana tabacum  51Nitrate reductase  49, 131Nitrite reductase  131Nuclear polyhedrosis viruses  74Nucleases  45, 119Nuclei  11, 78, 119Nucleocytoplasmic interaction  78, 99Nucleoproteins  83Nucleotide 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, 137Oncogenes  22, 62Organelles  77Ornithine carbamoyltransferase  20 Outbreaks  37Oxidoreductases  13, 14, 18, 20, 30, 53, 74, 79, 85, 94, 100Oxygen  30, 64Pathogenesis  10Pathogenesis-related proteins  70Pathogenicity  37Pentosyltransferases  83Peptides  44Peroxidases  73, 85Ph  38Phanerochaete chrysosporium  73, 74Phaseolus vulgaris  130Phenotypes  8, 84, 120, 126Phenylalanine ammonia-lyase  104Phormia terraenovae  44Phospholipase a2  111Phospholipids  117Phosphoproteins  50Phosphoric monoester hydrolases  19Phosphorylation  19Photosynthates  120Phytoalexins  94Phytochrome  82Phytophthora infestans  104, 129Phytotoxicity  48Pichia  88Pichia stipitis  14Pigeons  123Pisum sativum  54, 70Plant breeding  99Plant development  99Plant disease control  135Plant extracts  63Plant pathogenic fungi  91, 99, 129Plant pathogens  60Plant proteins  2, 5, 35, 70, 84, 96, 98, 102, 121, 133, 137Plant viruses  135Plasma membranes  5, 61, 69Plasmids  3, 47, 55, 66, 68, 69, 75, 86, 88, 100, 134Plasmodium falciparum  32Plastids  121Polygalacturonase  46Polymerase chain reaction  19, 80, 127Polypeptides  10Polyporus  85Potassium  16, 67Potyvirus group  57Precursors  63, 75Progesterone  3, 98Promoters  4, 20, 26, 32, 58, 64, 77, 87, 88, 98, 103, 113Protein composition  35Protein kinase  7, 50, 132Protein quality  109Protein secretion  38, 45, 111Protein synthesis  2, 33, 35, 40, 41, 89Proteinases  15, 48, 111Proteins  32, 55, 65, 88, 101, 108, 122, 126Proteolipids  136Proteolysis  29, 89, 102, 111Protoplasts  128Pseudocercosporella herpotrichoides  60Pseudogenes  63Purification  24, 74, 102, 112Pyruvate decarboxylase  103Pyruvate kinase  112Races  54Rats  43, 114Receptors  4Recombinant  DNA  4, 17, 18, 26, 32, 42, 55, 56, 69, 81, 96,98, 131, 134Recombinant vaccines  32Recombination  25, 38, 74, 77, 82, 112, 119Refeeding  133Regulation  5, 24Replication  65Reporter genes  26, 87, 100, 128, 129Resistance  48, 53, 84Restriction fragment length polymorphism  93Restriction mapping  17, 46, 63, 64, 103, 116, 121, 137Rhizopus  21, 59Rhodophyta  121Ribonucleases  58, 59Ribose  83Ribosomes  101Rna  48, 65, 84, 95, 127, 135Rna editing  63, 78Rna polymerase  58Rumen fungi  95Saccharomyces  28Saccharomyces 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, 137Saccharomyces uvarum  18Schizophyllum commune  126Screening  37, 95Secale cereale  60Secretory granules  61Segregation  8Semidominance  25Serine  50, 63Sesquiterpenes  51Sex pheromones  96Solanum tuberosum  104Somatostatin  75Soy protein  109Species differences  91Spectral analysis  49, 114Sporangia  33Spore germination  33Spores  33Sporulation  99Staphylococcus aureus  45Starch  26Starvation  133Steroid metabolism  3Strain differences  91Strains  12, 20, 58, 62, 91, 113, 117Stress response  70, 104Structural genes  6, 13, 24, 55, 64, 88, 103, 104, 115, 132,136 Structure activity relationships  72Substitution lines  99Substrates  76, 88Sugars  26Superoxide dismutase  1Suppression  106, 126Survival  39, 87Susceptibility  37Symptoms  37, 120Synergism  113Synthesis  28Synthetic genes  38, 123Targeted mutagenesis  48Temperature  105, 110Testosterone  3Thigmotropism  10Thunnus  123Tolerance  87Toxicity  66Transcription  1, 7, 22, 25, 27, 30, 31, 56, 58, 63, 64, 78,86, 95, 98, 101, 110, 113, 116, 121, 122, 127, 133Transfer  RNA  42, 63, 106Transferases  6, 47Transformation  69Transgenics  15, 51, 120Translation  5, 8, 20, 29, 89, 95, 121Translocation  34, 120Transport processes  61Transposable elements  11, 86, 116Triacylglycerol lipase  21Trichoderma  92Trifolium subterraneum  37Triticum aestivum  60, 99Triticum spelta  99Tubulin  91Tyrosine  19Uric acid  13Uromyces appendiculatus  10, 130Ustilago zeae  66Vacuoles  136Varietal susceptibility  54Vectors  3, 4, 9, 32, 34, 43, 55, 59, 66, 81, 88, 100, 114,123, 129, 131, 134Virulence  125Virus-like particles  66Western australia  37Wheat  52Wheat dwarf geminivirus  127Wheat germ  63Xylose  14Yeast extracts  63Yeasts  1, 20, 23, 41, 62, 67, 69, 76, 82, 105, 113, 118, 122Zea mays  71Zinc  1, 15