摘要:

AbstractUsing an improved procedure of pulsed field gel electrophoresis, yeast chromosomes were separated over a wide range of molecular size (250–4000 kbp) on single gels. The chromosomal DNA patterns of all the species belonging to the genusKluyveromyceswere examined. Within the speciesK. marxianus, the varietieslactis,drosophilarumandvanudeniishowed closely related patterns; very different from them, the varietiesbulgaricusandmarxianuswere related to each other, forming a distinct group; the strains commonly called ‘K. lactis’ and ‘K. fragilis’ were unambiguously different from each other in chromosome patterns. These differences were correlated with the presence of characteristic repetitive sequence elements in the mitochondrial DNA of the former group and not in the latter. Analysis ofCandida macedoniensis, which had been considered to be an anamorph ofK. marxianusvar.marxianus, showed that these two yeast species were indeed similar in chromosome patterns and in mitochondrial DNA restriction

ISSN:0749-503X    

DOI:10.1002/yea.320050103

出版商:John Wiley&Sons, Ltd.    

年代:1989

数据来源: WILEY

摘要:

AbstractTwo α‐glucosidase (maltase) genes, designatedGLUCPIandGLUCPII, have been cloned from an industrial strain of baker's yeast (Saccharomyces cerevisiae) by complementation of a maltase‐negative mutant strain. The different genes were identified according to their alternatively expressed isoenzymes PI and PII in transformants after isoelectric focusing and activity staining in separated cell lysates. The gene encoding α‐glucosidase PI (GLUCPI), which was not present in laboratory strains ofS. carlsbergensiswith a definedMAL1,2,3,4or6locus, was sequenced and compared with the recently publishedMAL6Sgene. This comparison revealed single amino acid deviations at three positions in the predicted polypeptide sequence. In addition, the divergent promoter region ofGLUCPIdiffered fromMAL6Sby a triple repeated 147‐bp DNA segment. Maltose induction and glucose repression of α‐glucosidase PI were not affected by the deletion of the repeated DNA segment. However, the absolute expression of α‐glucosidase PI increased two‐ to four‐fold. In addition, a two‐fold increase in the maltase synthesis occurred when the cloned positive regulator geneMAL2‐8cpwas on the same plasmid. Furthermore, stability of the α‐glucosidase in cultures in the stationary growth phase was greatly enhanced using a host strain lacking the proteinases A and B and the carboxypeptidases Y and S. Promoter trimming,MAL2‐8cpstimulation and the use of a host strain deficient in four vacuolar proteinases resulted in α‐glucosidase PI expression of a

ISSN:0749-503X    

DOI:10.1002/yea.320050104

出版商:John Wiley&Sons, Ltd.    

年代:1989

数据来源: WILEY

摘要:

AbstractThe KEX2 protease (product of theKEX2gene) functions late in the secretory pathway ofSaccharomyces cerevisiaeby cleaving the polypeptide chains of prepro‐killer toxin and prepro‐α‐factor at paired basic amino acid residues. The intracellular vesicles containing KEX2 protease sedimented in density gradients to a position distinct from those containing mannosyltransferase I (product of theMNN1gene), a marker enzyme for the Golgi complex. The recovery of intact compartments containing these enzymes approached 80% after sedimentation. We propose that the KEX2 protease and mannosyltransferase I reside within distinct compar

ISSN:0749-503X    

DOI:10.1002/yea.320050105

出版商:John Wiley&Sons, Ltd.    

年代:1989

数据来源: WILEY

摘要:

AbstractTheTRP1gene of the yeastKluyveromyces lactishas been cloned from a genomic library by complementation of theSaccharomyces cerevisiae trpl‐289 mutation. The gene was located within the clone by transposon mutagenesis and the coding region identified by DNA sequencing. This has indicated thatK. lactis TRP1encodes a 210‐amino acid polypeptide which shows 53% identity to the homologousS. cerevisiaeprotein. TheK. lactis TRP1gene has been disrupted by substituting theS. cerevisiae URA3gene for a large part of theTRP1coding sequence. Replacement of the chromosomalTRP1locus with this construction has enabled the production of non‐revertingtrp1−strains ofK. lactis, while a genetic analysis of the disrupted allele confirmed that theTRP1gene had been cloned. DNA sequencing has also shown that theK. lactis TRP1sequences is flanked by genes encoding inorganic pyrophosphatase and histone H3, which we have designatedIPPandHHT1respectively. Hybridization studies have shown that in common withS. cerevisiae,K. lactishas two copies of the histone H3 gene. Each H3 gene is closely linked to a gene encoding histone H4 and in both yeast species theIPPgene is tightly linked to one of the histone gen

ISSN:0749-503X    

DOI:10.1002/yea.320050106

出版商:John Wiley&Sons, Ltd.    

年代:1989

数据来源: WILEY

摘要:

AbstractMethods currently used for the extraction of proteins from yeast involve relatively long time periods between sampling cells from a culture and analysis of their proteins by polyacrylamide gel electrophoresis–sodium dodecylsulphate. Often it is desirable to inactivate cellular metabolism rapidly after sampling and here we show that trichloroacetic acid precipitation techniques, often used for rapid extraction and inactivation of proteins from higher eukaryotes, can be adapted for use with organisms which have cell wall

ISSN:0749-503X    

DOI:10.1002/yea.320050107

出版商:John Wiley&Sons, Ltd.    

年代:1989

数据来源: WILEY

摘要:

AbstractA newly isolated gene,ESS1, was shown to encode a protein required for vegetative growth inSaccharomyces cerevisiae. The nucleotide sequence ofESS1revealed a 172 amino acid open reading frame predicting a highly basic, 19·5 kilodalton product. Although the gene was isolated by cross‐hybridization with the vertebrate v‐sisoncogene, the primary amino acid sequence bears only a slight resemblance to the p28sisprotein.ESS1was shown to be single copy in the yeast genome and transcriptionally active during logarithmic growth. It is located on the right arm of chromosome X, 6 centimorgans distal toilv3. The genetic map location indicates it is not allelic to any previously characterized mutation in this organism. Both inactivation ofESS1by gene disruption and overexpression by fusion to a heterologous promoter were detrimental to growth in both haploid and diploid cell types. Under non‐permissive conditions, the terminal phenotype of strains containing a suppressible amber mutation withinESS1was one of aberrant multibudded structures. Examination of this morphology indicates that loss ofESS1function may lead to a defect in cytokinesis or cell sepa

ISSN:0749-503X    

DOI:10.1002/yea.320050108

出版商:John Wiley&Sons, Ltd.    

年代:1989

数据来源: WILEY

摘要:

AbstractThe effects of vandate on mitochondrial respiration and H+ATPase activity inSaccharomyces cerevisiaewere studied. A 50% inhibition of oxygen uptake in isolated mitochondria was produced by 4·4 mM‐V2O5. Activity of H+ATPase in whole mitochondria was inhibited by 50% by 5·5 μM‐V2O5, in submitochondrial particles by 55 μM‐V2O5; and in the chloroform‐released H+ATPase by 0·5 mM‐V2O5. Vandate was also found to relieve growth inhibition caused by the mitochondrial H+ATPase inhibitors NN′‐decyclohexylcarbodiimide and oligomycin. These results imply that vanadate could affect mitochondrial respiration by interacting with the H+ATPa

ISSN:0749-503X    

DOI:10.1002/yea.320050109

出版商:John Wiley&Sons, Ltd.    

年代:1989

数据来源: WILEY

ISSN:0749-503X    

DOI:10.1002/yea.320050102

出版商:John Wiley&Sons, Ltd.    

年代:1989

数据来源: WILEY