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  • AERODYNAMICS
  • Analytical Chemistry and Spectroscopy
  • Animals
  • Life and Medical Sciences
  • Physics
  • Yeast
  • Springer  (30)
  • 1990-1994  (30)
  • 1975-1979
  • 1925-1929
  • 1992  (30)
Collection
Keywords
Publisher
Years
  • 1990-1994  (30)
  • 1975-1979
  • 1925-1929
Year
  • 1
    Electronic Resource
    Electronic Resource
    Changes in protein composition ofSaccharomyces brewing strains in response to heat shock and ethanol stress (1992)
    Springer
    Journal of industrial microbiology and biotechnology 9 (1992), S. 229-234 
    Details
    ISSN: 1476-5535
    Keywords: Heat shock protein (HSP) ; Yeast ; Saccharomyces ; Viability ; Thermotolerance ; Ethanol tolerance
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Summary Heat shock and ethanol stress of brewing yeast strains resulted in the induction of a set of proteins referred to as heat shock proteins (HSPs). At least six strongly induced HSPs were identified in a lager brewing strain and four HSPs in an ale brewing strain. Four of these HSPs with molecular masses of approximately 70, 38, 26 and 23 kDa were also identified in two laboratory strains ofSaccharomyces cerevisiae. The appearance of HSPs correlated with increased survival of strains at elevated temperatures and high concentrations of ethanol. These results suggest that HSPs may play a role in the ethanol and thermotolerance of yeasts. The properties of these proteins and membrane fatty acids in relation to heat and ethanol shock are being investigated.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF01569628
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  • 2
    Electronic Resource
    Electronic Resource
    Genetic analysis of ubiquitin-dependent protein degradation (1992)
    Springer
    Cellular and molecular life sciences 48 (1992), S. 172-178 
    Details
    ISSN: 1420-9071
    Keywords: Yeast ; protein degradation ; ubiquitin conjugating enzymes ; signals for proteolysis ; stress response
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract Selective degradation of cellular proteins serves to eliminate abnormal proteins and to mediate the turnover of certain short-lived proteins, many of which have regulatory functions. In eukaryotes a major pathway for selective protein degradation is ATP-dependent and is mediated by the ubiquitin system. This pathway involves substrate recognition by components of a ubiquitin-protein ligase system, covalent attachment of ubiquitin moieties to proteolytic substrates, and subsequent degradation of these conjugates by a multicatalytic protease complex. Recent genetic evidence suggests that the remarkable selectivity of this process is largely controlled at the level of substrate recognition by the ubiquitin ligase system. InSaccharomyces cerevisiae, ubiquitin-conjugating enzymes UBC1, UBC4 and UBC5 have been identified as key components of this highly conserved degradation pathway. Genetic analysis indicates that ubiquitin-dependent proteolysis is essential for cell viability and that UBC4 and UBC5 enzymes are essential components of the eukaryotic stress response.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF01923510
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  • 3
    Electronic Resource
    Electronic Resource
    The unusual reversion properties of a mitochondrial mutation in the structural gene of subunit I of cytochrome oxidase of Saccharomyces cerevisiae reveal a probable histidine ligand of the redox center (1992)
    Springer
    Current genetics 21 (1992), S. 147-151 
    Details
    ISSN: 1432-0983
    Keywords: Yeast ; Mitochondria ; Cytochrome oxidase
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary We have analyzed a mutation in the mitochondrial gene oxi3 coding for subunit I of cytochrome-oxidase in the yeast Saccharomyces cerevisiae. This mutation replaces one of the seven invariant histidines of the polypeptide (position 378) by a tyrosine, and leads to a respiratory deficient phenotype. A total of 157 revertants, which have recovered the ability to grow on a respiratory substrate, have been selected from this mutant (tyrosine 378). The nature of the reversion has been analysed by a rapid screening procedure and 32 of the revertants have been sequenced. They are all true backmutations reintroducing the histidine in position 378. This very exceptional situation suggests that this histidine is a ligand of the redox center of cytochrome oxidase.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00318474
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  • 4
    Electronic Resource
    Electronic Resource
    Positive cis-acting regulatory sequences mediate proper control of POL1 transcription in Saccharomyces cerevisiae (1992)
    Springer
    Current genetics 21 (1992), S. 183-189 
    Details
    ISSN: 1432-0983
    Keywords: Yeast ; DNA-polymerase α ; Cell cycle
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary The 5′ ACGCGT3′ MluI motif, which is found in the upstream region of several yeast DNA-synthesis genes which are periodically expressed during the mitotic cell-cycle, is present twice in the 5′ non-coding region of the DNA-polymerase α gene (POL1). Deletion, of the most distal repeat does not affect POL1 transcription, while the adjacent 40 base-pair (bp) downstream sequence is necessary both for the proper level and the fluctuation of POL1 mRNA. This region contains the 5′ACGCGTCGCGT3′ sequence, which is sufficient to control periodic transcription of a CYC1-lacZ reporter gene with the same kinetics observed for POL1. The adjacent 29 bp AT-rich region does not show any activity by itself, but it acts synergistically in conjunction with at least one MluI hexamer to stimulate CYC1-lacZ expression. By further deletion analysis, DNA sequences necessary to initiate POL1 transcription at the proper sites have also been identified.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00336839
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  • 5
    Electronic Resource
    Electronic Resource
    Direct selection of galactokinase-negative mutants of Candida albicans using 2-deoxy-galactose (1992)
    Springer
    Current genetics 21 (1992), S. 203-206 
    Details
    ISSN: 1432-0983
    Keywords: Yeast ; Galactokinase ; Mutant selection
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary The galactose analogue 2-deoxy-galactose (2DG) has been widely used to select for mutations in the gene encoding the galactose pathway enzyme galactokinase (GalK). We have tested the effect of 2DG on Candida albicans to see if it could be used to obtain GalK- mutants in this diploid asexual yeast. 2DG was shown to be toxic to wild-type cells. Enzyme assays demonstrated that 2DG can induce GalK as efficiently as galactose. Examination of the initital rate of galactose uptake indicated that the galactose transport system is constitutive. 2DG-resistant mutants were isolated from mutagenized cultures and shown to have very low levels of GalK activity. The potential genetic applications of this system of direct mutant selection are discussed.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00336842
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  • 6
    Electronic Resource
    Electronic Resource
    The UGA43 negative regulatory gene of Saccharomyces cerevisiae contains both a GATA-1 type zinc finger and a putative leucine zipper (1992)
    Springer
    Current genetics 21 (1992), S. 301-307 
    Details
    ISSN: 1432-0983
    Keywords: Repressor ; Zinc finger ; Leucine zipper ; GATA-1 ; Yeast
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary The UGA43 gene of Saccharomyces cerevisiae is required for repression of inducible genes involved in the utilization of 4-aminobutyric acid (GABA) or urea as nitrogen sources. The UGA43 gene has been cloned by complementation of a uga43 mutation. The N-terminal region of the UGA43 protein is very similar to the DNA-binding zinc-finger region typical of the GATA regulatory factor family in vertebrates. UGA43 is the first reported instance of a GATA protein with a negative regulatory function. The C-terminal region of the predicted UGA43 protein contains a putative leucine zipper. Sequencing of three uga43 mutant alleles suggests that the GATA and putative leucine-zipper regions are both required for the repressive activity of UGA43. UGA43 appears to be a highly regulated gene. On “poor” nitrogen sources, UGA43 transcripts are measured at high levels whereas they are nearly undetectable in conditions of nitrogen catabolite repression. The levels measured on “poor” nitrogen sources are further increased in uga43 mutant cells, suggesting that UGA43 exerts negative autoregulation.
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    URL: http://dx.doi.org/10.1007/BF00351687
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  • 7
    Electronic Resource
    Electronic Resource
    The CCS1 gene from Saccharomyces cerevisiae which is involved in mitochondrial functions is identified as IRA2 an attenuator of RAS1 and RAS2 gene products (1992)
    Springer
    Current genetics 21 (1992), S. 325-329 
    Details
    ISSN: 1432-0983
    Keywords: Yeast ; cAMP ; RAS ; GAP
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary The ccs1-1 mutation of Saccharomyces cerevisiae, which has been previously described, is associated with an increase in cytochrome content, in respiration, and in ATP synthesis. In addition, this mutation leads to the same phenotype as cells de-regulated in the cAMP pathway. From a yeast genomic library, we have isolated a DNA fragment in a recombinant plasmid pCD1 which complements the ccs1-1 mutation. Homologous integration of this DNA in the genome occurs at the CCS1 locus. An 11 kb of the DNA insert is necessary for complementation. Sequencing part of the fragment identifies CCS1 as the IRA2 gene. The IRA2 gene is known to encode an attenuator of RAS gene product activity which stimulates the GTPase activity of the RAS proteins. This result underlines the involvement of cAMP-dependent phosphorylation in mitochondrial function. We present the sequence of 1 kb DNA upstream of the putative ATG of the IRA2/CCS1 gene product which is devoid of an ORF and could contain several regulatory sites.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00351690
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  • 8
    Electronic Resource
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    In vitro mutagenesis of the mitochondrial leucyl-tRNA synthetase of S. cerevisiae reveals residues critical for its in vivo activities (1992)
    Springer
    Current genetics 22 (1992), S. 69-74 
    Details
    ISSN: 1432-0983
    Keywords: Yeast ; Mitochondria ; Aminoacyl-tRNA synthetase ; RNA splicing
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary The mitochondrial leucyl-tRNA synthetase (mLRS) of Saccharomyces cerevisiae is involved in both mitochondrial protein synthesis and pre-mRNA splicing. We have created mutations in the regions HIGH, GWD and KMSKS, which are involved in ATP-, amino acid-and tRNA-binding respectively, and which have been conserved in the evolution of group I tRNA synthetases. The mutants GRD and NMSKS have no discernible phenotype. The mutants AWD and ARD act as null alleles and lead to the production of 100% cytoplasmic petites. The mutants HIGN, NIGH and KMSNS are unable to grown on glycerol even in the presence of an intronless mitochondrial genome and accumulate petites to a greater extent than the wild-type but less than 40%. Experiments with an imported bI4 maturase indicate that the lesion in these mutations primarily affects the synthetase and not the splicing functions.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00351744
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  • 9
    Electronic Resource
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    Comparative analysis of spontaneous mitotic recombination in [cir 0] and [cir +] strains of the yeast Saccharomyces cerevisiae (1992)
    Springer
    Current genetics 22 (1992), S. 259-265 
    Details
    ISSN: 1432-0983
    Keywords: Yeast ; 2 μm plasmid ; Mitotic recombination ; Coincident conversion
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary The influence of the 2 μm plasmid on homologous recombination in the right arm of chromosome XV of the yeast Saccharomyces cerevisiae has been examined. No differences between spontaneous mitotic recombination rates in [cir 0] and [cir +] derivatives of two yeast diploid tester strains were detected. In the course of analysis an unusually high coincident conversion frequency at ADE2, HIS3, and two RFLP loci adjacent to ADE2, was observed. The character of coincident homozygotization of linked markers argues for a “break-and-replicate” mechanism underlying the coincident conversion events.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00317918
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  • 10
    Electronic Resource
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    The REV2 gene of Saccharomyces cerevisiae: cloning and DNA sequence (1992)
    Springer
    Current genetics 22 (1992), S. 277-282 
    Details
    ISSN: 1432-0983
    Keywords: Yeast ; DNA-repair ; Mutation-deficient mutant ; Nucleotide-binding consensus
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary The REV2 gene of Saccharomyces cerevisiae was cloned and sequenced; it contains an open reading frame of 1985 bp with a coding potential of 662 amino acids. Interruption of the chromosomal REV2 gene by integrating the URA3 gene coupled with partial deletion of the 3′ terminal region produced viable haploid rev2Δ mutants. This indicates that the REV2 gene is non-essential for growth. The rev2Δ mutant is slightly more UV-sensitive than strains carrying various rev2 alleles (rev2-1, rev2x, rad5-1, rad5-8). The putative Rev2 protein is probably a globular protein containing a highly conserved nucleotide-binding site and two zinc-finger domains.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00317921
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  • 11
    Electronic Resource
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    A ten-minute protocol for transforming Saccharomyces cerevisiae by electroporation (1992)
    Springer
    Current genetics 22 (1992), S. 335-336 
    Details
    ISSN: 1432-0983
    Keywords: Yeast ; Rapid transformation ; Cell age
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary We present a simplified and rapid method for the transformation of yeast cells by electroporation. Stationary cells, scraped off the agar of Petri dish cultures stored in the refrigerator for up to 6 weeks, are suspended in sorbitol buffer, spun down by gentle centrifugation, transferred into the electroporation cuvette, and immediately subjected to transformation via electroporation. Transformation efficiency of this 10-min method, which does not require the preparation of cell cultures, is about 10% of the hitherto best performing transformation procedure using cells of defined growth phase.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00317931
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  • 12
    Electronic Resource
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    Regulation of mitochondrial transcription during the stringent response in yeast (1992)
    Springer
    Current genetics 21 (1992), S. 241-247 
    Details
    ISSN: 1432-0983
    Keywords: Yeast ; Transcription ; Mitochondria ; RNA
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary In yeast (S. cerevisiae) the stringent response is known to include rapid, selective, and severe transcriptional curtailment for genes specifying cytoplasmic rRNAs and r-proteins. We have shown that transcription of the mitochondrial 21S rRNA gene is also congruently and selectively curtailed during the yeast stringent response. Using an in vitro transcription assay with intact organelles from both ϱ+ and ϱ− strains, we show here that the mitochondrial stringent response includes not only transcription of the 21S and 16S rRNA genes, but also that of organellar genes specifying non-mitoribosome-related products. Stringent organellar transcriptional curtailment is identical when cells are starved for a required (marker) amino acid or when they are subjected to nutritional downshift, and the relative level of that transcriptional curtailment following either perturbation is the same in cells growing on fermentative (repressing) or purely respiratory carbon sources. These results confirm that the mechanism governing mitochondrial gene expression during a stringent response is specified outside the organelle, and they show that this transcriptional control mechanism is not immediately subject to glucose repression. In all strains examined, stringent organellar gene expression requires a mitochondrial promoter, suggesting that the regulatory mechanism which functions during the stringent response operates primarily at transcriptional initiation.
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    URL: http://dx.doi.org/10.1007/BF00336848
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  • 13
    Electronic Resource
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    The gene for cadmium metallothionein from a cadmium-resistant yeast appears to be identical to CUP 1 in a copper-resistant strain (1992)
    Springer
    Current genetics 21 (1992), S. 275-280 
    Details
    ISSN: 1432-0983
    Keywords: Yeast ; Cadmium resistance ; Metallothionein gene ; CUP 1
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary A cadmium-resistant strain of Saccharomyces cerevisiae produces a cadmium metallothionein with the same characteristics as the copper metallothionein that is encoded by CUP 1 in a copper-resistant strain. The structural gene for metallothionein from the cadmium-resistant strain resembles CUP 1 in terms of the fragmentation patterns generated by restriction enzymes. Furthermore, the gene may be amplified as 2.0 kb repeating units in both the cadmium-resistant and the copperresistant strains. However, transformants with a plasmid that carried the metallothionein gene from the cadmiumresistant strain were resistant to copper but not to cadmium. It appears that the same metallothionein gene, CUP 1, is amplified in both cadmium- and copper-resistant yeasts. However, the mechanism for the cadmiumspecific inducibility of the gene may be restricted to the cadmium-resistant strain.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00351682
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  • 14
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    Interaction of the yeast pleiotropic drug resistance genes PDR1 and PDR5 (1992)
    Springer
    Current genetics 21 (1992), S. 431-436 
    Details
    ISSN: 1432-0983
    Keywords: Drug resistance ; Yeast ; Positive activator
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary The network of genes which mediates multiple drug resistance in yeast includes, among others, the PDR1 gene, which encodes a putative regulator of gene expression, and PDR5, a locus whose amplification leads to resistance. We demonstrate that disruption of PDR5 causes marked hypersensitivity not only to cycloheximide but also to sulphometuron methyl and the mitochondrial inhibitors chloramphenicol, lincomycin, erythromycin and antimycin. Genetic analysis of double mutants containing an insertion in PDR5 (pdr5:Tn5), which renders cells hypersensitive to cycloheximide, and a pdr1 mutation, which confers resistance to this inhibitor, indicates that the expression of resistance requires a functional PDR5 gene. The same interdependency is observed for chloramphenicol, but not for oligomycin, lincomycin, crythromycin or sulphometuron methyl. Northern analysis of PDR1 and PDR5 transcripts reveals that the 5.2 kbp PDR5 transcript is overexpressed in pdr1 (resistant) mutants, but underexpressed in a disruption of PDR1. These observations provide strong experimental support for our former proposal that the PDR5 gene is a target for regulation by the PDR1 gene product.
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    URL: http://dx.doi.org/10.1007/BF00351651
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  • 15
    Electronic Resource
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    One-step transformation of yeast in stationary phase (1992)
    Springer
    Current genetics 21 (1992), S. 83-84 
    Details
    ISSN: 1432-0983
    Keywords: Yeast ; Transformation ; Thio compound ; Stationary phase
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary A fast yeast-transformation technique has been developed by adding thio compounds to alkali-ion based protocols and incubating at 45°C. This procedure is especially recommended for cells from stationary phase at a density up to 2.5×108 cells/ml. It involves only one step for the preparation and transformation of competent cells within 30 min. The yield was more than 104 transformants/μg plasmid DNA. This protocol is easy to scale up for many DNA samples and is also applicable for yeast cells from different types of storages.
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    URL: http://dx.doi.org/10.1007/BF00318659
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  • 16
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    A promoter deletion reduces the rate of mitotic, but not meiotic, recombination at the HIS4 locus in yeast (1992)
    Springer
    Current genetics 21 (1992), S. 109-116 
    Details
    ISSN: 1432-0983
    Keywords: Transcription ; Recombination ; Yeast ; Gene conversion
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Several investigators have reported that transcription stimulates some types of mitotic recombination in the yeast Saccharomyces cerevisiae. We find that mutations that reduce the rate of trancription of the yeast HIS4 gene in vegetative cells reduce the frequency of mitotic, but not meiotic, recombination events.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00318468
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  • 17
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    Mitotic hyperploidy for chromosomes VIII and III in Saccharomyces cerevisiae (1992)
    Springer
    Current genetics 21 (1992), S. 309-318 
    Details
    ISSN: 1432-0983
    Keywords: Yeast ; Aneuploidy ; Chromosomes
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary The arg4–8 and cup1 s markers comprise a copy-number-dependent signal device in the yeast Saccharomyces cerevisiae. These alleles permit reliable discrimination between euploid and disomic haploids as well as between euploid and trisomic diploids. To investigate and compare inherent inter-chromosomal differences as regards propensity for hyperploidy, we transplaced arg4–8 and cup1 s by deleting them from chromosome VIII and then re-introducing them at the leu2 locus on chromosome III. The rate of chromosome gain was significantly greater for the chromosome III construct compared to the native chromosome VIII, in both diploid and haploid strains. In addition, more coincident aneuploidy for other chromosomes was found among chromosome VIII hyperploids compared to chromosome III hyperploids.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00351688
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  • 18
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    Complementation of the Saccharomyces cerevisiae srb1-1 mutation: an autoselection system for stable plasmid maintenance (1992)
    Springer
    Current genetics 21 (1992), S. 339-344 
    Details
    ISSN: 1432-0983
    Keywords: Yeast ; Saccharomyces cerevisiae ; Lysis mutants ; Plasmid stability
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary The autonomously replicating plasmid YEpSS1, containing the S. cerevisiae SOD1 and SRB1 genes, was highly unstable in a wild-type strain. When transformed into a fragile srb1-1 mutant host, the same plasmid displayed different characteristics depending on the growth medium used. Both batch and continuous culture experiments demonstrated that the plasmid was very unstable when the transformed strain SLU15 was grown in the presence of an osmotic stabiliser (10% w/v sorbitol). However, in the absence of the osmoticum, nearly 100% of the cells retained the plasmid and produced the Sod1 protein after 80 generations of growth.
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    URL: http://dx.doi.org/10.1007/BF00351692
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  • 19
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    Adaptation and major chromosomal changes in populations of Saccharomyces cerevisiae (1992)
    Springer
    Current genetics 22 (1992), S. 13-19 
    Details
    ISSN: 1432-0983
    Keywords: Chromosome length variants ; Adaptation ; Yeast ; Continuous culture
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Thirteen independent populations of Saccharomyces cerevisiae (nine haploid and four diploid) were maintained in continuous culture for up to approximately 1000 generations, with growth limited by the concentration of organic phosphates in medium buffered at pH 6. Analysis of clones isolated from these populations showed that a number (17) of large-scale chromosomallength variants and rearrangements were present in the populations at their termination. Nine of the 16 yeast chromosomes were involved in such changes. Few of the changes could be explained by copy-number increases in the structural loci for acid phosphatase. Several considerations concerning the nature and frequency of the chromosome-length variants observed lead us to conclude that they are selectively advantageous.
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    URL: http://dx.doi.org/10.1007/BF00351736
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  • 20
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    Mutational analysis of a variant of ARS1 from Saccharomyces cerevisiae (1992)
    Springer
    Current genetics 22 (1992), S. 175-180 
    Details
    ISSN: 1432-0983
    Keywords: ARS1 mutants ; DNA replication ; Yeast ; Single-stranded DNA
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary A naturally occurring single base-pair G to A transition, creating a 10/11 near-match close to the essential 11 base-pair core consensus of ARS1, was used to investigate the importance of near-match sequences. The 10/11 near-match can not substitute for the core consensus since an ARS- phenotype is observed when the core consensus is deleted. However, deletion mutations revealed that this near-match together with a short palindromic sequence, also situated in the B-flanking region, comprise a single element crucial for optimal ARS function. The palindrome has the potential of forming a stemloop structure. Rather precise observations concerning the borders of the B-region were achieved. The four base pairs separating the near-match from the core consensus perform a spacing function where the identity of the bases are unimportant. However, this spacing is highly important since deletion of these four base pairs leads to an ARS- phenotype.
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    URL: http://dx.doi.org/10.1007/BF00351723
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  • 21
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    Effects of heat shock and ethanol stress on the viability of aSaccharomyces uvarum (carlsbergensis) brewing yeast strain during fermentation of high gravity wort (1992)
    Springer
    Journal of industrial microbiology and biotechnology 10 (1992), S. 111-116 
    Details
    ISSN: 1476-5535
    Keywords: Heat shock ; Ethanol ; Saccharomyces ; Yeast ; Fermentation ; Viability ; Wort
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Summary The effects of heat shock and ethanol stress on the viability of a lager brewing yeast strain during fermentation of high gravity wort were studied. These stress effects resulted in reduced cell viability and inhibition of cell growth during fermentation. Cells were observed to be less tolerant to heat shock during the fermentation of 25°P (degree Plato) wort than cells fermenting 16°P wort. Degree Plato (oP) is the weight of extract (sugar) equivalent to the weight of sucrose in a 100 g solution at 20°C. Relieving the stress effects of ethanol by washing the cells free of culture medium, improved their tolerance to heat shock. Cellular changes in yeast protein composition were observed after 24 h of fermentation at which time more than 2% (v/v) ethanol was present in the growth medium. The synthesis of these proteins was either induced by ethanol or was the result of the transition of cells from exponential phase to stationary phase of growth. No differences were observed in the protein composition of cells fermenting 16°P wort compared to those fermenting 25°P wort. Thus, the differences in the tolerance of these cells to heat shock may be due to the higher ethanol concentration produced in 25°P wort which enhanced their sensitivity to heat shock.
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    URL: http://dx.doi.org/10.1007/BF01583843
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  • 22
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    A new member of the adenylate kinase family in yeast: PAK3 is highly homologous to mammalian AK3 and is targeted to mitochondria (1992)
    Springer
    Molecular genetics and genomics 233 (1992), S. 363-371 
    Details
    ISSN: 1617-4623
    Keywords: Adenylate kinase family ; Yeast ; Mitochondria
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Making use of the polymerase chain reaction primed by oligonucleotides corresponding to regions conserved between members of the nucleoside monophosphate kinase family, we have isolated the yeast gene PAK3. Pak3p belongs to the subgroup of long-form adenylate kinase isozymes (deduced molecular mass 25.3 kDa) and exhibits highest sequence similarity to bovine AK3 rather than to the yeast isozyme, Aky2p. The gene is shown to be non-essential because haploid disruption mutants are viable, both in the presence and absence of a functional AKY2 allele. It maps on chromosome V upstream of RAD3. Its expression level is low when cells are grown on glucose or other fermentable carbon sources and about threefold higher on glycerol, but can be significantly induced by ethanol. A PAK3/mouse dihydrofolate reductase fusion construct expressed in yeast is targeted to mitochondria. Transformation with PAK3 on a multicopy plasmid complements neither adenylate kinase deficiency in an aky2-disrupted yeast strain nor in Escherichia coli cells conditionally defective in adenylate kinase.
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    URL: http://dx.doi.org/10.1007/BF00265432
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    Molecular cloning of the imidazoleglycerolphosphate dehydratase gene of Trichoderma harzianum by genetic complementation in Saccharomyces cerevisiae using a direct expression vector (1992)
    Springer
    Molecular genetics and genomics 234 (1992), S. 481-488 
    Details
    ISSN: 1617-4623
    Keywords: Filamentous fungi ; Complementation ; Yeast ; Histidine biosynthetic genes ; Shuttle vector
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary The Trichoderma harzianum imidazoleglycerolphosphate dehydratase gene (igh) has been isolated by complementation of a Saccharomyces cerevisiae his3 mutant using a direct expression vector. This Escherichia coli-yeast shuttle vector was developed to allow efficient cloning and expression of cDNA libraries. The cDNA is 627 nucleotides long and codes for a protein of 209 amino acids with an apparent molecular mass of 22 466 daltons. The predicted protein sequence showed 63.6%, 58.7%, and 38.4% identity respectively to the corresponding enzymes from S. cerevisiae, Pichia pastoris and E. coli. Northern analysis showed that the expression of the igh gene in T. harzianum is not inhibited by external histidine and the level of igh mRNA was about threefold higher in cells starved of histidine.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00538709
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    Targeting sequences of the two major peroxisomal proteins in the methylotrophic yeast Hansenula polymorpha (1992)
    Springer
    Molecular genetics and genomics 235 (1992), S. 269-278 
    Details
    ISSN: 1617-4623
    Keywords: Peroxisomes ; Targeting signals ; Yeast ; Hansenula polymorpha
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Dihydroxyacetone synthase (DAS) and methanol oxidase (MOX) are the major enzyme constituents of the peroxisomal matrix in the methylotrophic yeast Hansenula polymorpha when grown on methanol as a sole carbon source. In order to characterize their topogenic signals the localization of truncated polypeptides and hybrid proteins was analysed in transformed yeast cells by subcellular fractionation and electron microscopy. The C-terminal part of DAS, when fused to the bacterial β-lactamase or mouse dihydrofolate reductase, directed these hybrid polypeptides to the peroxisome compartment. The targeting signal was further delimited to the extreme C-terminus, comprising the sequence N-K-L-COOH, similar to the recently identified and widely distributed peroxisomal targeting signal (PTS) S-K-L-COOH in firefly luciferase. By an identical approach, the extreme C-terminus of MOX, comprising the tripeptide A-R-F-COOH, was shown to be the PTS of this protein. Furthermore, on fusion of a C-terminal sequence from firefly luciferase including the PTS, β-lactamase was also imported into the peroxisomes of H. polymorpha. We conclude that, besides the conserved PTS (or described variants), other amino acid sequences with this function have evolved in nature.
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    URL: http://dx.doi.org/10.1007/BF00279370
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    STE50, a novel gene required for activation of conjugation at an early step in mating in Saccharomyces cerevisiae (1992)
    Springer
    Molecular genetics and genomics 236 (1992), S. 145-154 
    Details
    ISSN: 1617-4623
    Keywords: Cell differentiation ; G protein ; Adaptation ; STE50 ; Yeast
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary A new gene, STE50, which plays an essential role in cell differentiation in Saccharomyces cerevisiae was detected and analysed. STE50 expression is not cell type-specific and its expression in MAT a and MATα cells is unaffected by pheromones. When present on a high copy number plasmid, STE50 causes supersensitivity to α-pheromone, and increases the level of α-pheromone-induced transcription of FUS1 in haploid a cells. Mutants bearing either of the two gene disruptions, ste50-1 or ste50-2, are sterile and have a modulated sensitivity to α-pheromone. The overexpression of STE4 (Gβ) in wild-type cells elicits a constitutive growth arrest signal, however this phenotype is suppressed by a C-terminal truncation mutation in STE50 (ste50-2). In contrast, the constitutive activation of the pheromone response pathway caused by disruption of GPA1 (Gα) is not suppressed in ste50-2 mutants. The ste50-2 mutation partially suppresses the desensitisation defect of the sst2-1 mutation, and the resulting ste50-2 sst2-1 mutants restore fertility. Our result sindicate that the ste50-2 mutant may have a defect in adaptation (hyperadaptation), and suggest a possible interaction of STE50-2 with the Gα subunit of the G protein.
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    URL: http://dx.doi.org/10.1007/BF00279653
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    Structure and regulation of yeast HEM3, the gene for porphobilinogen deaminase (1992)
    Springer
    Molecular genetics and genomics 234 (1992), S. 233-243 
    Details
    ISSN: 1617-4623
    Keywords: Yeast ; Heme ; Sequence ; Regulation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Porphobilinogen deaminase is the third enzyme in the heme biosynthetic pathway. hem3 mutants in Saccharomyces cerevisiae are deficient in porphobilinogen deaminase activity. We have isolated the HEM3 gene by complementation of the heme auxotrophy of a hem3 mutant. Sequence analysis reveals an open reading frame of 981 nucleotides. The derived amino acid sequence of the protein encoded by HEM3 shows extensive homology to the reported sequences for porphobilinogen deaminase from a number of other sources, indicating that HEM3 is the structural gene for porphobilinogen deaminase. Earlier reports have suggested that expression of HEM3 is induced by porphobilinogen, the substrate of the encoded enzyme. We have investigated the transcription of HEM3 and have found that it is not affected by the ability of the cell to make porphobilinogen or heme. However, we have found that HAP2 and HAP3 gene products are involved in the expression of HEM3. An important element required for expression of HEM3 has been localized to a small region that contains a sequence homologous to the HAP2-3-4 binding sites of several genes including HEM1. These findings suggest that HEM3 expression is regulated in the same manner as that of HEM1 which encodes the first enzyme of the heme biosynthetic pathway.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00283844
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    Effect of donor copy number on the rate of gene conversion in the yeast Saccharomyces cerevisiae (1992)
    Springer
    Molecular genetics and genomics 235 (1992), S. 97-103 
    Details
    ISSN: 1617-4623
    Keywords: Gene conversion ; Yeast ; Plasmid ; Recombination
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Nonreciprocal recombination (gene conversion) between homologous sequences at nonhomologous locations in the genome occurs readily in the yeast Saccharomyces cerevisiae. In order to test whether the rate of gene conversion is dependent on the number of homologous copies available in the cell to act as donors of information, the level of conversion of a defined allele was measured in strains carrying plasmids containing homologous sequences. The level of recombination was elevated in a strain carrying multiple copies of the plasmid, compared with the same strain carrying a single copy of the homologous sequences either on a plasmid or integrated in the genome. Thus, the level of conversion is proportional to the number of copies of donor sequences present in the cell. We discuss these results within the framework of currently favoured models of recombination.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00286186
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    Isolation and phenotypic analysis of conditional-lethal, linker-insertion mutations in the gene encoding the largest subunit of RNA polymerase II in Saccharomyces cerevisme (1992)
    Springer
    Molecular genetics and genomics 232 (1992), S. 408-414 
    Details
    ISSN: 1617-4623
    Keywords: Linker-insertion mutagenesis ; RNA polymerase II ; Conditional-lethal mutants ; Yeast
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Linker-insertion mutagenesis was used to isolate mutations in the Saccharomyces cerevisiae gene encoding the largest subunit of RNA polymerase II (RP021, also called RPBI). The mutant rpo21 alleles carried on a plamid were introduced into a haploid yeast strain that conditionally expresses RP021 from the inducible promoter pGAL10. Growth of this strain on medium containing glucose is sustained only if the plasmid-borne rpo21 allele encodes a functional protein. Of nineteen linker-insertion alleles tested, five (rpo21-4 to −8) were found that impose a temperature-sensitive (ts) lethal phenotype on yeast cells. Four of these five is alleles encode mutant proteins in which the site of insertion lies near one of the regions of the largest subunit that have been conserved during evolution. Two of the is mutants (rpo21-4 and rpo21-7) display pleiotropic phenotypes, including an auxotrophy for inositol and a decreased proliferation rate at the permissive temperature. The functional relationship between RP021 and RP026, the gene encoding the 17.9 kDa subunit shared by RNA polymerases 1, 11, and III was investigated by determining the ability of increased dosage of RP026 to suppress the is phenotype imposed by rpo21-4 to −8. Suppression of the is defect was specific for the rpo21-4 allele and was accompanied by co-suppression of the inositol auxotrophy. These results suggest that mutations in the largest subunit of RNA polymerase II can have profound effects on the expression of specific subsets of genes, such as those involved in the metabolism of inositol. In the rpo21-4 mutant, these pleiotropic phenotypes can be attributed to a defective interaction between the largest subunit and the RP026 subunit of RNA polymerase II.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00266244
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    TheSaccharomyces cerevisiae GAM2/SIN3 protein plays a role in both activation and repression of transcription (1992)
    Springer
    Molecular genetics and genomics 233 (1992), S. 327-330 
    Details
    ISSN: 1617-4623
    Keywords: GAM2 ; SIN3 ; Transcriptional regulator ; Glucoamylase ; Yeast
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary We have cloned GAM2, which is required for transcription ofSTA1, a gene encoding an extracellular glucoamylase inSaccharomyces cerevisiae var.diastaticus. DNA sequence analysis revealed thatGAM2 is the same gene asSIN3, known to be a general negative regulator of yeast genes. RNA blot analysis indicated thatGAM2/SIN3 also acts as a positive regulator ofGAM3/ADR6, which in turn is required for transcription ofSTA1 andADH2. These results suggest thatGAM2 regulatesSTA1 expression through transcriptional activation ofGAM3 and indicate that GAM2/SIN3 protein is a transcriptional regulator that can play a role in both activation and repression of transcription.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00587597
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    An alignment of 17 deduced protein sequences from plant, fungi, and ciliate H+-ATPase genes (1992)
    Springer
    Journal of bioenergetics and biomembranes 24 (1992), S. 309-317 
    Details
    ISSN: 1573-6881
    Keywords: Yeast ; proton ATPase ; sequence alignment ; hydrophobic regions
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology , Physics
    Notes: Abstract Seventeen protein sequences of H+-ATPases from plants (Arabidopsis thaliana, Nicotiana plumbaginifolia, Lycopersicum esculentum), fungi (Sacharomyces cerevisiae, Schizosaccharomyces pombe, Zygosaccharomyces rouxii, Neuropora crassa, Candida albicans), and a parasitic ciliate (Leishmania donovani) have been aligned. Twenty sequence fragments were identified which were conserved in H+-, Na+/K+-, and Ca++ plasma membrane-ATPases. In addition, a total of 118 residues not located in these fragments were found to be conserved in all H+-ATPases. Among those, 38 amino acid residues were screened out as being priority targets for site-directed mutagenesis experiments aimed at the identification of the amino acid residues specifically involved in cation specificity.
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    URL: http://dx.doi.org/10.1007/BF00768851
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