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  • Yeast  (35)
  • AERODYNAMICS
  • Analytical Chemistry and Spectroscopy
  • Animals
  • Life and Medical Sciences
  • Physics
  • Springer  (35)
  • 1990-1994  (35)
  • 1975-1979
  • 1925-1929
  • 1990  (35)
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Keywords
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  • 1990-1994  (35)
  • 1975-1979
  • 1925-1929
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  • 1
    Electronic Resource
    Electronic Resource
    Fermentation of hardwood hemicellulose hydrolysate byPachysolen tannophilus, candida shehatae andPichia stipitis (1990)
    Springer
    Journal of industrial microbiology and biotechnology 6 (1990), S. 157-164 
    Details
    ISSN: 1476-5535
    Keywords: Lignocellulosic waste ; Yeast ; Ethanol production ; Optimization study
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Summary Hardwood hemicellulose hydrolysate has been utilized as a substrate for ethanol production. Among the three different yeasts tested, the best performances have been obtained, in decreasing order, usingPachysolen tannophilus, Candida shehatae andPichia stipitis. Several pretreatments of this raw material have been studied to improve ethanol yields; in one such pretreatment a strain ofP. tannophilus produced ethanol with a yield of 0.29 gethanol/gsugars (gP/gS); which is only 15% less than the values observed with synthetic media. Neither aeration nor acetone addition improved the fermentation of this substrate; in fact, only a marked stimulation of biomass growth has been observed at the expense of both ethanol and xylitol production.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF01577690
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  • 2
    Electronic Resource
    Electronic Resource
    The intron of the yeast actin gene contains the promoter for an antisense RNA (1990)
    Springer
    Current genetics 17 (1990), S. 269-273 
    Details
    ISSN: 1432-0983
    Keywords: Yeast ; Actin ; Intron ; Antisense RNA
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Using Northern blot analysis we have detected an approximately 840 nucleotide-long RNA which is complementary to the 5′ leader sequence and the first ten nucleotides of the coding sequence of the yeast actin (ACT1) messenger RNA. We have determined two transcription start sites for this actin antisense RNA (ASR1), both within the ACT1 intron, at about 80 and 90 nucleotides downstream from the 5′ splice site. Analysis of a cDNA clone showed that this RNA species overlaps the entire trailer sequence and approximately 20 nucleotides of the coding sequence of the nearby yeast YPT1 gene.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00312620
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  • 3
    Electronic Resource
    Electronic Resource
    Mitochondrial modifications in a single nuclear mutant of Saccharomyces cerevisiae affected in cAMP-dependent protein phosphorylation (1990)
    Springer
    Current genetics 17 (1990), S. 507-513 
    Details
    ISSN: 1432-0983
    Keywords: Yeast ; Mutants ; Cytochrome ; Mitochondria ; Oxidative phosphorylation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary This paper reports studies of bioenergetic modifications in a TTR1 single-nuclear mutant, isolated as resistant to triethyltin, an inhibitor of mitochondrial ATPase, and effective in cAMP-dependent protein phosphorylation. This mutant appears to have lost the wildtype cell ability to respond to a decrease of oxygen concentration in the growth medium by a decrease of cytochrome concentration in the cell. ATP synthesis rate in mutant cells in both the prestationary and stationary phase of growth appeared increased in comparison to wild-type cells, as too was respiration rate. A comparative study of mitochondria extracted from wild-type and from TTR1 mutant cells showed an increase in respiration rate, an increase in ATP synthesis rate, and an increase in TPP+ uptake in mutant mitochondria. The specific ATPase activity, as well as its sensitivity to TET, appears to be similar for mitochondria extracted from both strains. It was proposed that the modification of mitochondrial biogenesis in the TTR1 mutant may be due to a response of the cell to an increase in ATP hydrolysis caused by the mutation. It is also possible that the modification in cAMP-dependent protein kinase regulation which appeared to occur in this mutant affects protein(s) involved in mitochondrial biogenesis.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00313079
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  • 4
    Electronic Resource
    Electronic Resource
    Chromosome location of a family of genes encoding different acidic ribosomal proteins in Saccharomyces cerevisiae (1990)
    Springer
    Current genetics 17 (1990), S. 535-536 
    Details
    ISSN: 1432-0983
    Keywords: Yeast ; Chromosome mapping ; Acidic ribosomal proteins
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary DNA probes from the genes encoding the acidic ribosomal proteins L44, L44′ and L45, as well as from reporter genes for chromosomes IV, VII, XII and XV, have been hybridised to Southern blots of Saccharomyces cerevisiae DNA resolved by pulsed field gel electrophoresis. The protein L44′ and protein L45 genes have been found to hybridise to chromosome IV, identified by the CAT1 gene probe, while the protein L44 probe hybridises with a band containing chromosomes VII and XV, identified by the ATPase 1 and HIS3 genes respectively.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00313084
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  • 5
    Electronic Resource
    Electronic Resource
    Yeast ribosomal proteins: XI. Molecular analysis of two genes encoding YL41, an extremely small and basic ribosomal protein, from Saccharomyces cerevisiae (1990)
    Springer
    Current genetics 17 (1990), S. 185-190 
    Details
    ISSN: 1432-0983
    Keywords: Yeast ; Ribosomal protein gene ; Sequence analysis
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Two genes encoding ribosomal protein YL41 were cloned from Saccharomyces cerevisiae chromosomal DNA. Both genes contain an uniterrupted region of only 75 nucleotides coding for a protein of 3.3 kD. Within the coding regions the nucleotide sequences are virtually identical, whereas in both the 5′-and 3′-flanking regions the two genes differ significantly from each other. The deduced protein shows an arginine and lysine content of 68 percent, i.e., 17 out of 25 residues, and the basic residues are evenly distributed over the molecule. When compared to the ribosomal protein sequences currently available no counterpart to YL41 could be found in prokaryotes and it seems likely that YL41 is a eukaryotespecific ribosomal protein.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00312608
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  • 6
    Electronic Resource
    Electronic Resource
    The DNA-binding protein RAP1 is required for efficient transcriptional activation of the yeast PYK glycolytic gene (1990)
    Springer
    Current genetics 18 (1990), S. 405-412 
    Details
    ISSN: 1432-0983
    Keywords: Yeast ; Trans-acting Factor ; RAP1
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary We show by deletion mutagenesis, followed by in vivo and in vitro analysis, that the binding of a protein factor to the upstream activation sequence (USA) of the Saccharomyces cerevisiae glycolytic gene PYK, encoding pyruvate kinase, is required for efficient transcription of the corresponding coding region. In addition, gel electrophoretic mobility shift and DNase I protection studies, involving yeast gene products expressed in E. coli, suggest that this trans-acting DNA-binding protein is encoding by the RAP1 gene. The identification of RAP1 binding sites located within the UAS element of the yeast PYK, PGK (phosphoglycerate kinase) and ENO1 (enolase) genes, and in the 5′-upstream region of the ADHI (alcohol dehydrogenase) gene, suggests that a mechanism of coordinate gene expression involving several of the glycolytic genes may exist in yeast.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00309909
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  • 7
    Electronic Resource
    Electronic Resource
    Ty virus-like particles in the Saccharomyces cerevisiae strain NCYC74 (1990)
    Springer
    Current genetics 18 (1990), S. 485-491 
    Details
    ISSN: 1432-0983
    Keywords: Yeast ; Ty elements ; Virus like particles
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Electron microscopic analysis of thin sections of Saccharomyces cerevisiae NCYC74 has revealed the presence of many clumped cytoplasmic particles that morphologically resemble Ty element virus-like particles (VLPs). Accumulation of Ty VLPs has only previously been observed in S. cerevisiae strains that over-express a cloned Ty element. The particles in NCYC74 co-purify with Ty RNA, Ty-specific antigens and a reverse transcriptase activity. Furthermore, they appear to be recognised by antibodies to Ty VLPs during indirect immunofluorescence experiments. These observations provide compelling evidence that the cytoplasmic particle in NCYC74 are indeed Ty VLPs.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00327018
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  • 8
    Electronic Resource
    Electronic Resource
    Analysis of interchromosomal mitotic recombination (1990)
    Springer
    Current genetics 18 (1990), S. 29-39 
    Details
    ISSN: 1432-0983
    Keywords: Recombination ; DNA repair ; UV irradiation ; Yeast
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary A novel synthetic locus is described that provides a simple assay system for characterizing mitotic recombinants. The locus consists of the TRP1 and HIS3 genes inserted into chromosome III of S. cerevisiae between the CRY1 and MAT loci. Defined trp1 and his3 alleles have been generated that allow the selection of interchromosomal recombinants in this interval. Trp+ or His+ recombinants can be divided into several classes based on coupling of the other alleles in the interval. The tight linkage of the CRY1 and MAT loci, combined with the drug resistance and cell type phenotypes that they respectively control, facilitates the classification of the recombinants without resorting to tetrad dissection. We present the distribution of spontaneous recombinants among the classes defined by this analysis. The data suggest that the recombination intermediate can have regions of symmetric strand exchange and that co-conversion tracts can extend over 1–3 kb. Continuous conversion tracts are favored over discontinuous tracts. The distribution among the classes defined by this analysis is altered in recombinants induced by UV irradiation.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00321112
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  • 9
    Electronic Resource
    Electronic Resource
    Two group I mitochondrial introns in the cob-box and coxI genes require the same MRS1/PET157 nuclear gene product for splicing (1990)
    Springer
    Current genetics 18 (1990), S. 117-124 
    Details
    ISSN: 1432-0983
    Keywords: Yeast ; Mitochondrial RNA splicing ; Nuclear pet - mutant ; Group I introns
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary We have studied the role of the product of the nuclear gene PET157 in mitochondrial pre-mRNA splicing. Cytoduction experiments show that a mitochondrial genome deleted for the three introns bI3, aI5 and aI6 is able to suppress the pet157-1 mutation: the strain recovers respiratory competency indicating that the product of the PET157 gene is only required for mitochondrial premRNA splicing. Characterization of the high molecular weight pre-mRNAs which accumulate in the pet157 mutant demonstrate that the product of the PET157 gene is required for the excision of two group I introns bI3 and aI6 (corresponding to aI5β) located in the cob-box and coxI genes respectively. Furthermore, the pet157 mutant strain accumulates the bI3 maturase in the form of a polypeptide of 50K (p50) previously observed in mitochondrial mutants defective in the excision of bI3. We have shown by restriction analysis and allelism tests that the pet157-1 mutation is allelic to the nuclear mrs1 mutation, previously described as specifically blocking the excision of bI3. Finally, revertants obtained by the deletion of bI3 or aI6 from the mitochondrial DNA were isolated from the MRS1 disrupted allele, confirming the involvment of the product of the MRS1/PET157 gene in the excision of the two introns bI3 and aI6.
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    URL: http://dx.doi.org/10.1007/BF00312599
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  • 10
    Electronic Resource
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    Resistance to cadmium is under the control of the CAD2 gene in the yeast Saccharomyces cerevisiae (1990)
    Springer
    Current genetics 18 (1990), S. 181-185 
    Details
    ISSN: 1432-0983
    Keywords: S. cerevisiae ; Yeast ; Cadmium resistance ; CAD2 gene
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary A cadmium-resistant strain, X3382-3A, which is able to grow in a medium containing 0.2 mM cadmium sulfate, was picked out from our laboratory stock strains of Saccharomyces cerevisiae. The cadmium resistance of this strain is controlled by a single dominant nuclear gene, denoted as CAD2. The locus of CAD2 was mapped by gene linkage to a site 15.5 centimorgans to the right of the his7 locus on the right arm of chromosome II. The cadmium resistance of the strain carrying CAD2 was evaluated for its properties of cadmium uptake, cadmium distribution and cadmium-metallothionein formation, in comparison with those of some other strains. The results suggest that the novel type of cadmium resistance controlled by CAD2 does not involve production of a cadmiumm-metallothionein.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00318377
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  • 11
    Electronic Resource
    Electronic Resource
    Effects of hap mutations on heme and cytochrome formation in yeast (1990)
    Springer
    Current genetics 17 (1990), S. 179-183 
    Details
    ISSN: 1432-0983
    Keywords: Heme ; Cytochromes ; Regulation ; Yeast
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Simultaneous effects of mutations in the transcriptional regulatory genes, HAP1, HAP2 and HAP3, on all respiratory cytochromes of Saccharomyces cerevisiae were determined. Cytochrome behavior in hap mutants and in cyc4 and rhm1 mutants, altered in regulation of 5-aminolevulinate synthase, was compared. Although hap mutants were isolated as trans-acting, transcriptional regulators of the CYC1 (iso-1-cytochrome c) gene, each mutant exhibits partial deficiencies in all cytochrome types. In hap2 and hap3 strains all cytochromes were decreased proportionally to about 40–50% of wild type values. In contrast, hap1 caused a decrease in all cytochromes and an accumulation of a pigment, probably Zn porphyrin. Apparently apocytochrome and heme biosynthesis retain coordination in hap2 and hap3, but not in hap1, mutants. Unlike cyc4 and rhm1 mutants, hap mutants do not exhibit 5-aminolevulinate-dependent restoration of cytochromes. The hap1 mutant grew at nearnormal rates on glycerol, whereas hap2 and hap3 mutants grew very slowly. The frequency of [rho-] was high (16–18%) in hap2 and hap3 strains. Results are consistent with generalized control of mitochondrial replication directed by the HAP1-HAP2 system and heme-directed control of formation of all apocytochromes mediated by HAP1. Neither system exerts all-or-nothing control.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00312865
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  • 12
    Electronic Resource
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    The effect of DNA replication on mutation of the Saccharomyces cerevisiae CDC8 gene (1990)
    Springer
    Current genetics 17 (1990), S. 275-280 
    Details
    ISSN: 1432-0983
    Keywords: Yeast ; DNA replication ; Effect on mutation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Incubation in YPD medium under permissive conditions when DNA replication is going on, strongly stimulates the induction of cdc+ colonies of UV-irradiated cells of yeast strains HB23 (cdc8-1/cdc8-3), HB26 (cdc8-3/cdc8-3) and HB7 (cdc8-1/cdc8-1). Inhibition of DNA replication by hydroxyurea, araCMP, cycloheximide or caffeine or else by incubation in phosphate buffer pH 7.0, abolishes this stimulation. Thus the replication of DNA is strongly correlated with the high induction of cdc+ colonies by UV irradiation. It is postulated that these UV-induced cdc+ colonies arise as the result infidelity in DNA replication.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00314872
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  • 13
    Electronic Resource
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    The yeast Yarrowia lipolytica has two, functional, signal recognition particle 7S RNA genes (1990)
    Springer
    Current genetics 17 (1990), S. 289-292 
    Details
    ISSN: 1432-0983
    Keywords: Yarrowia lipolytica ; 7SL RNA ; Essential genes ; Yeast
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Cells containing a deletion of either the SCR1 or SCR2 genes, which code for the 7SL RNA component of the signal recognition particle (SRP) homologue, were found to be viable. Two independent approaches demonstrated that cells containing deletions of both genes were inviale. Therefore, Yarrowia lipolytica contains two (and only two) functional 7SL RNA genes.
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    URL: http://dx.doi.org/10.1007/BF00314874
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  • 14
    Electronic Resource
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    The RAD50 gene, a member of the double strand break repair epistasis group, is not required for spontaneous mitotic recombination in yeast (1990)
    Springer
    Current genetics 18 (1990), S. 111-116 
    Details
    ISSN: 1432-0983
    Keywords: Mitotic recombination ; Hyper-recombination ; RAD50 ; Yeast
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Mutations in the RAD50 gene of Saccharomyces cerevisiae have been shown to reduce double strand break repair, meiotic recombination, and radiation-inducible mitotic recombination. Several different point mutations (including ochre and amber alleles) have been previously examined for effects on spontaneous mitotic recombination and did not reduce the frequency of recombination. Instead, the rad50 mutations conferred a moderate hyper-rec phenotype. This paper examines a deletion/interruption allele of RAD50 that removes 998 of 1312 amino acids and adds 1.1 kb of foreign DNA. The results clearly indicate that spontaneous mitotic recombination can occur in the absence of RAD50; in fact, the frequency of recombination is elevated over the wild-type cell. One possible interpretation of these observations is that the initiating lesion in spontaneous recombination events in mitosis might not be a double strand break.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00312598
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  • 15
    Electronic Resource
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    Flow cytometry as a tool to discriminate respiratory-competent and respiratory-deficient yeast cells (1990)
    Springer
    Current genetics 18 (1990), S. 265-267 
    Details
    ISSN: 1432-0983
    Keywords: Flow cytometry ; Rhodamine 123 ; Respiratory chain ; Yeast
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary The cationic lipophilic dye Rhodamine 123 (Rh123) is selectively enriched in mitochondria in a membrane potential-dependent manner. Application of drugs which interfere with the electron flow of the respiratory chain lead to a severe reduction of mitochondrial dye uptake. In this communication we show that the same effect is observed after Rh123-staining of respiratory-deficient yeast mutants. Based on this observation we used flow cytometry to discriminate respiratory-compentent and respiratory-deficient yeast cells. Combined with a cell sorter we were able to selectively enrich respiring and non-respiring yeast cells, repectively, from a mixture of cells.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00318391
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  • 16
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    Expression of the gene encoding subunit II of yeast QH2: Cytochrome c oxidoreductase is regulated by multiple factors (1990)
    Springer
    Current genetics 17 (1990), S. 459-464 
    Details
    ISSN: 1432-0983
    Keywords: Yeast ; QH2: cytochrome c oxidoreductase ; Mitochondrial biogenesis ; Transcription
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary In Saccharmmyces cerevisiae, the COR2 gene codes for the 40 kDa subunit II of the QH2: cytochrome c oxidoreductase, an enzyme of the mitochondrial respiratory chain. Regions in the 5′ flank of this gene important for regulated expression were identified by assaying β-galactosidase activities in cells carrying different COR2-lacZ fusion genes. Sequences downstream of position-201 relative to the translational initiation codon are sufficient to confer regulation by carbon source, whereas sequences downstream of position-153 do not give rise to significant expression. A binding site for the abundant general transcription factor GFI is present in the region between-201 and-153 just upstream from sequences which resemble the consensus DNA recognition sequence of the regulatory protein complex HAP2/HAP3: 5′-TNATTGGT-3′. By quantitating RNA levels and assaying β-galactosidase activities we show that synthesis of COR2, which is not a hemoprotein, is regulated by HAP1, HAP2/HAP3 and heme.
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    URL: http://dx.doi.org/10.1007/BF00313072
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  • 17
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    Isolation and genetic study of Triethyltin-resistant mutants of Saccharomyces cerevisiae (1990)
    Springer
    Current genetics 17 (1990), S. 465-472 
    Details
    ISSN: 1432-0983
    Keywords: Yeast ; Mutant ; Triethyltin chloride ; Protein phosphorylation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Three mutants of Saccharomyces cerevisiae resistant to triethyltin (an inhibitor of mitochondrial ATPase) on non-fermentative media, and non-resistant to this drug on fermentative media, were isolated and named TTR1, TTR2 and TTR3. Apart from triethyltin resistance, these mutants show the following common characteristics: (1) Increased intracellular cytochrome c concentration. (2) Increased respiration rate. (3) Decreased growth yield. (4) Increased growth sensitivity to several drugs inhibiting oxidative phosphorylation: namely, CCCP (permeabilizing inner mitochondrial membrane to protons), valinomycin (permeabilizing inner mitochondrial membrane to potassium) and oligomycin (inhibitor of mitochondrial ATPase). (5) Increased sensitivity to carbon source starvation. For each mutant, these characteristics appeared to be due to a single pleiotropic nuclear mutation. Mutation TTR1 causes additional phenotypic characteristics which do not appear in mutants TTR2 and TTR3: (1) Pinkish coloration of colonies which is more pronounced after a long growth period. (2) Inability of the cells to store glycogen. (3) Growth defect of the cells on a galactose-containing medium. (4) Inability of a diploid homozygote mutant strain to sporulate. All these phenotypic characteristics have already been described in yeast mutants deregulated in cAMP-dependant protein phosphorylation. Crossing of a strain bearing the TTR1 mutation with a strain mutated in the adenylate cyclase structural gene suggested that the TTR1 phenotype is due to a modification in regulation of cAPK by cAMP, making cell multiplication possible without intracellular cAMP.
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    URL: http://dx.doi.org/10.1007/BF00313073
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  • 18
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    Characterization of a respiratory-deficient mutant of Pachysolen tannophilus (1990)
    Springer
    Current genetics 17 (1990), S. 493-497 
    Details
    ISSN: 1432-0983
    Keywords: Mitochondria ; Yeast ; Petites
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary A pleiotropic, respiration-deficient mutant was isolated from the petite negative yeast Pachysolen tannophilus after UV mutagenesis. The mutant is unable to utilize xylose, arabinose, galactose or glycerol, and shows no detectable respiration when grown on glucose. Cytochrome c oxidase, xylose reductase and xylitol dehydrogenase activities are lacking. Mitochondrial ultrastructre is altered. The results support the hypothesis that functioning mitochondria are necessary for xylose utilization in this organism.
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    URL: http://dx.doi.org/10.1007/BF00313077
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  • 19
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    Blockage to exonuclease III digestion in the chromatin of Saccharomyces cerevisiae maps to the in vitro — determined binding site of a trans-acting regulatory factor (1990)
    Springer
    Current genetics 18 (1990), S. 17-22 
    Details
    ISSN: 1432-0983
    Keywords: Yeast ; Ty2 ; Protein/DNA binding ; Transcription
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary A series of transposable element-induced mutations at the HIS4 locus in Saccharomyces cerevisiae have been attributed to the transposition of a Ty element into the 5′ regulatory region of this gene. Various Ty-containing His+ revertants have been isolated and the HIS4/Ty junction region sequenced. The only difference found in this region between a His- and a weak His+ strain was a single point mutation, an A→G transition. The position of Ty remained unaltered. Examination of lacZ fusion plasmids further implicated this A→G transition as being reponsible for the altered phenotype, the bp transition representing an allele of a cis-acting regulatory element. Subsequent gel retardation and methylation interference experiments revealed that this A→G mutation enabled the binding of a trans-acting factor (TyBf) in vitro. In this paper we show that the TyBf binding site is in a region of chromatin hypersensitive to digestion by DNase I. The binding site is protected in vivo from digestion with exonuclease III, suggesting the presence of a bound protein in His+ (“on”) but not His- (“off”) Ty-containing strains. We propose that a trans-acting factor binding in vivo, presumably TyBf, is responsible for the activation of HIS4 expression in these insertion mutants.
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    URL: http://dx.doi.org/10.1007/BF00321110
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  • 20
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    Yeast SCO1 protein is required for a post-translational step in the accumulation of mitochondrial cytochrome c oxidase subunits I and II (1990)
    Springer
    Current genetics 18 (1990), S. 13-15 
    Details
    ISSN: 1432-0983
    Keywords: Yeast ; Mitochondria ; Cytochrome c oxidase ; Post-translational regulation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Biogenesis of functional cytochrome c oxidase in yeast requires the product of the nuclear gene SCO1. Strains deleted for this gene fail to accumulate the mitochondrially-synthesized cytochrome c oxidase subunits I and II, despite the presence of the respective mRNAs. Here we present data which demonstrate that the observed phenotype does not result from a failure to translate the mRNAs, but from a preferential degradation of the newly synthesized subunits. The SCO1 protein is therefore involved in a post-translational step in the accumulation of cytochrome c oxidase subunits I and II. We propose that the SCO1 protein is required for the correct assembly of both subunits into the cytochrome c oxidase complex.
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    URL: http://dx.doi.org/10.1007/BF00321109
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  • 21
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    Proliferation and metabolic significance of peroxisomes in Candida boidinii during growth on d-alanine or oleic acid as the sole carbon source (1990)
    Springer
    Archives of microbiology 153 (1990), S. 485-489 
    Details
    ISSN: 1432-072X
    Keywords: Candida boidinii ; Yeast ; Peroxisomes ; β-Oxidation ; d-Amino acid oxidase
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract We have studied the induction of peroxisomes in the methylotrophic yeast Candida boidinii by d-alanine and oleic acid. The organism was able to utilize each of these compounds as the sole carbon source and grew with growth rates of μ=0.20 h-1 (on d-alanine) or μ=0.43 h-1 (on oleic acid). Growth was associated with the development of many peroxisomes in the cells. On d-alanine a cluster of tightly interwoven organelles was observed which made up 6.3% of the cytoplasmic volume and were characterized by the presence of d-amino acid oxidase and catalase. On oleic acid rounded to elongated peroxisomes were dominant which were scattered throughout the cytoplasm. These organelles contained increased levels of β-oxidation enzymes; their relative volume fraction amounted 12.8% of the cytoplasmic volume.
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    URL: http://dx.doi.org/10.1007/BF00248431
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  • 22
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    Purification and characterization of a (R)-2,3-butanediol dehydrogenase from Saccharomyces cerevisiae (1990)
    Springer
    Archives of microbiology 154 (1990), S. 267-273 
    Details
    ISSN: 1432-072X
    Keywords: Yeast ; Saccharomyces cerevisiae ; (R)-2,3-Butanediol dehydrogenase ; Stereospecificity ; Gas chromatographic analysis of enantiomers
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract A NAD-dependent (R)-2,3-butanediol dehydrogenase (EC 1.1.1.4), selectively catalyzing the oxidation at the (R)-center of 2,3-butanediol irrespective of the absolute configuration of the other carbinol center, was isolated from cell extracts of the yeast Saccharomyces cerevisiae. Purification was achieved by means of streptomycin sulfate treatment, Sephadex G-25 filtration, DEAE-Sepharose CL-6B chromatography, affinity chromatography on Matrex Gel Blue A and Superose 6 prep grade chromatography leading to a 70-fold enrichment of the specific activity with 44% yield. Analysis of chiral products was carried out by gas chromatographic methods via pre-chromatographic derivatization and resolution of corresponding diasteromeric derivatives. The enzyme was capable to reduce irreversibly diacetyl (2,3-butanediol) to (R)-acetoin (3-hydroxy-2-butanone) and in a subsequent reaction reversibly to (R,R)-2,3-butanediol using NADH as coenzyme. 1-Hydroxy-2-ketones and C5-acyloins were also accepted as substrates, whereas the enzyme was inactive towards the reduction of acetone and dihydroxyacetone. The relative molecular mass (M r) of the enzyme was estimated as 140 000 by means of gel filtration. On SDS-polyacrylamide gel the protein decomposed into 4 (identical) subunits of M r 35 000. Optimum pH was 6.7 for the reduction of acetoin to 2,3-butanediol and 7.2 for the reverse reaction.
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    URL: http://dx.doi.org/10.1007/BF00248966
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    Killer toxin of Hanseniaspora uvarum (1990)
    Springer
    Archives of microbiology 154 (1990), S. 175-178 
    Details
    ISSN: 1432-072X
    Keywords: Killer toxin ; Hanseniaspora uvarum ; Yeast
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The yeast Hanseniaspora uvarum liberates a killer toxin lethal to sensitive strains of the species Saccharomyces cerevisiae. Secretion of this killer toxin was inhibited by tunicamycin, an inhibitor of N-glycosylation, although the mature killer protein did not show any detectable carbohydrate structures. Culture supernatants of the killer strain were concentrated by ultrafiltration and the extracellular killer toxin was precipitated with ethanol and purified by ion exchange chromatography. SDS-PAGE of the electrophoretically homogenous killer protein indicated an apparent molecular mass of 18,000. Additional investigations of the primary toxin binding sites within the cell wall of sensitive yeast strains showed that the killer toxin of Hanseniaspora uvarum is bound by β-1, 6-d-glucans.
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    URL: http://dx.doi.org/10.1007/BF00423329
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  • 24
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    13C NMR analysis of production and accumulation of osmoregulatory metabolites in the salt-tolerant yeast Debaryomyces hansenii (1990)
    Springer
    Archives of microbiology 154 (1990), S. 209-214 
    Details
    ISSN: 1432-072X
    Keywords: 13C NMR spectroscopy ; Yeast ; Debaryomyces hansenii ; Osmoregulation ; Compatible solute ; Salt stress
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract High resolution 13C NMR combined with chemical analysis were used to study the formation of metabolites from [1-13C]-labelled glucose by the salt-tolerant yeast Debaryomyces hansenii after transfer to media containing 8% NaCl. Time course spectroscopy of an aerobic cell suspension showed [1,3-13C]glycerol as the predominant end product. Perchloric acid extracts revealed additional less prominent incorporation of label into arabinitol, trehalose, glutamic acid, and alanine. The incorporation into trehalose and arabinitol showed a transient increase after shift to the high salinity medium. It is concluded that glycerol and arabinitol are the major organic solutes in D. hansenii, the production of glycerol being strongly induced by high salinity. Analysis of labelled extracts of D. hansenii after transfer to 8% NaCl media containing [1-13C]- or [6-13C]glucose, demonstrated that glucose is dissimilated via a combination of the Embden-Meyerhof-Parnas pathway and the pentose phosphate pathway, with the former playing a major role in glycerol formation and the latter in arabinitol production. The almost exclusive labelling of C5 of arabinitol from [6-13C]glucose indicates that the pathway to arabinitol proceeds via reduction of ribulose-5-phosphate.
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    URL: http://dx.doi.org/10.1007/BF00248956
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  • 25
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    Absence of glucose-induced cAMP signaling in the Saccharomyces cerevisiae mutants cat1 and cat3 which are deficient in derepression of glucose-repressible proteins (1990)
    Springer
    Archives of microbiology 154 (1990), S. 199-205 
    Details
    ISSN: 1432-072X
    Keywords: cAMP ; Cat mutants ; Glucose repression ; Glucose-induced ; Intracellular pH ; Ras ; Saccharomyces cerevisiae ; Signal transduction ; Trehalase ; Yeast
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Addition of glucose to derepressed cells of the yeast Saccharomyces cerevisiae induces a transient, specific cAMP signal. Intracellular acidification in these cells, as caused by addition of protonophores like 2,4-dinitrophenol (DNP) causes a large, lasting increase in the cAMP level. The effect of glucose and DNP was investigated in glucose-repressed wild type cells and in cells of two mutants which are deficient in derepression of glucose-repressible proteins, cat1 and cat3. Addition of glucose to cells of the cat3 mutant caused a transient increase in the cAMP level whereas cells of the cat1 mutant and in most cases also repressed wild type cells did not respond to glucose addition with a cAMP increase. The glucose-induced cAMP increase in cat3 cells and the cAMP increase occasionally present in repressed wild type cells however could be prevented completely by addition of a very low level of glucose in advance. In derepressed wild type cells this does not prevent the specific glucose-induced cAMP signal at all. These results indicate that repressed cells do not show a true glucose-induced cAMP signal. When DNP was added to glucose-repressed wild type cells or to cells of the cat1 and cat3 mutants no cAMP increase was observed. Addition of a very low level of glucose before the DNP restored the cAMP increase which points to lack of ATP as the cause for the absence of the DNP effect. These data show that intracellular acidification is able to enhance the cAMP level in repressed cells. The glucose-induced artefactual increase occasionally observed in repressed cells is probably caused by the fact that their low intracellular pH is only restored after the ATP level has increased to such an extent that it is no longer limiting for cAMP synthesis. It is unclear why the artefactual increases are not always observed. Measurement of glucose- and DNP-induced activation of trehalase confirmed the physiological validity of the changes observed in the cAMP level. Our results are consistent with the idea that the glucose-induced signaling pathway contains a glucose-repressible protein and that the protein is located before the point where intracellular acidification triggers activation of the pathway.
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    URL: http://dx.doi.org/10.1007/BF00423333
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  • 26
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    Expression of the yeast DNA primase gene, PRI1, is regulated within the mitotic cell cycle and in meiosis (1990)
    Springer
    Molecular genetics and genomics 221 (1990), S. 44-48 
    Details
    ISSN: 1617-4623
    Keywords: Yeast ; Cell cycle ; Meiosis ; DNA primase
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Mitotic cultures synchronised either by a feed-starve protocol or by elutriation have been used to show that the Saccharomyces cerevisiae DNA primase I gene is periodically expressed in the cell cycle. The transcript increases many-fold in late G1 and reaches a peak at the same time as four other genes essential for DNA synthesis, CDC8, CDC9, CDC21 and POL1. The primase I transcript is also regulated in meiosis, reaching maximal levels during premeiotic DNA synthesis.
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    URL: http://dx.doi.org/10.1007/BF00280366
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  • 27
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    Initiation of meiosis and sporulation in Saccharomyces cerevisiae requires a novel protein kinase homologue (1990)
    Springer
    Molecular genetics and genomics 221 (1990), S. 176-186 
    Details
    ISSN: 1617-4623
    Keywords: Meiosis ; Sporulation ; Northern hybridization ; Regulatory circuit ; Yeast
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary SME1 was cloned due to its high copy number effect: it enabled MATα/MATα diploid cells to undergo meiosis and sporulation in a vegetative medium. Disruption of SME1 resulted in a recessive Spo− phenotype. These results suggest that SME1 is a positive regulator for meiosis. DNA sequencing analysis revealed an open reading frame of 645 amino acids. An amino terminal peptide of ca 400 amino acids in the deduced protein was similar to known protein kinases. Transcription of SME1 was regulated negatively by nitrogen and glucose and positively by MATα/MATα and IME1, another positive regulator gene of meiosis. By complementation analysis, SME1 was found to be identical to IME2, which had been shown to be important in meiosis. These results suggest that IME1 product stimulates meiosis by activating transcription of SME1 (IME2) and that protein phosphorylation is required for initiation of meiosis.
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    URL: http://dx.doi.org/10.1007/BF00261718
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  • 28
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    The yeast mitochondrial leucyl-tRNA synthetase is a splicing factor for the excision of several group I introns (1990)
    Springer
    Molecular genetics and genomics 224 (1990), S. 209-221 
    Details
    ISSN: 1617-4623
    Keywords: Aminoacyl-tRNA synthetase ; RNA splicing ; Group I introns ; RNA maturase ; Yeast
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary The Saccharomyces cerevisiae nuclear gene NAM2 codes for mitochondrial leucyl-tRNA synthetase (mLRS). Herbert et al. (1988, EMBO J 7:473–483) proposed that this protein is involved in mitochondrial RNA splicing. Here we present the construction and analyses of nine mutations obtained by creating two-codon insertions within the NAM2 gene. Three of these prevent respiration while maintaining the mitochondrial genome. These three mutants: (1) display in vitro a mLRS activity ranging from 0%–50% that of the wild type: (2) allow in vivo the synthesis of several mitochondrially encoded proteins; (3) prevent the synthesis of the COXII protein but not of its mRNA; (4) abolish the splicing of the group I introns bI4 and aI4; and (5) affect significantly the excision of the group I introns bI2, bI3 and aI3. Importation of the bI4 maturase from the cytoplasm into mitochondria in a nam2 − mutant strain does not restore the excision of the introns bI4 and aI4 implying that the splicing deficiency does not result from the absence of the bI4 maturase. We conclude that the mLRS is a splicing factor essential for the excision of the group I introns bI4 and aI4 and probably important for the excision of other group I introns.
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    URL: http://dx.doi.org/10.1007/BF00271554
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    TheSaccharomyces cerevisiae NPR1 gene required for the activity of ammonia-sensitive amino acid permeases encodes a protein kinase homologue (1990)
    Springer
    Molecular genetics and genomics 222 (1990), S. 393-399 
    Details
    ISSN: 1617-4623
    Keywords: General amino acid permease ; Protein kinase ; Serine-rich protein ; Transport protein ; Yeast
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary TheNPR1 gene ofSaccharomyces cerevisiae plays a central role in controlling permease activity; its product is required to promote the activity of at least six distinct transport systems for nitrogenous nutrients under conditions of nitrogen catabolite derepression. We report here the nucleotide sequence of the clonedNPR1 gene. The predicted amino acid sequence indicates thatNPR1 encodes a protein of 86 kDa which appears to be organized into two distinct structural domains. The amino-terminal domain of NPR1 (residues 1 to 440) contains 26% serine residues and several regions strongly enriched for PEST residues suggesting a short half-life for the NPR1 protein. The carboxy-terminal region of NPR1 contains consensus sequences characteristic of the catalytic domains of protein kinases. Therefore, NPR1-dependent positive control of nitrogen transport systems most likely involves protein phosphorylation. Northern analysis indicates that the absence of general amino acid permease (GAP1) activity innpr1 mutants is not due to reduction in transcription or messenger stability. Hence, the NPR1 protein probably acts at the post-transcriptional level. Proteins that may serve as substrates for phosphorylation are discussed.
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    URL: http://dx.doi.org/10.1007/BF00633845
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    Functional analysis of ARGRI and ARGRIII regulatory proteins involved in the regulation of arginine metabolism inSaccharomyces cerevisiae (1990)
    Springer
    Molecular genetics and genomics 222 (1990), S. 192-200 
    Details
    ISSN: 1617-4623
    Keywords: Yeast ; Arginine ; Regulatory protein
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary We present here a functional analysis of ARGRI and ARGRIII regulatory proteins which are involved together with ARGRII in specific regulation of arginine anabolic and catabolic pathways. Unlike ARGRII, ARGRI and ARGRIII have no transcriptional activation capacity. The first 60 amino acids of ARGRI (out of 177) are dispensable for its activity. The functional domain of the protein is located in the region of homology with MCM1 and SRF proteins. ARGRIII contains in its C-terminal portion a stretch of 17 aspartate residues which are indispensable for arginine regulation. Gene disruption of theARGRIII gene impairs the growth of the mutant on rich medium, showing that ARGRIII has a pleiotropic role in the cell.
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    URL: http://dx.doi.org/10.1007/BF00633817
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    Magnification of the rDNA cluster in Kluyveromyces lactis (1990)
    Springer
    Molecular genetics and genomics 223 (1990), S. 342-344 
    Details
    ISSN: 1617-4623
    Keywords: rRNA genes ; Growth rate ; Yeast ; Pulsed field gel electrophoresis
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary By employing pulsed field gel electrophoresis we find that slow growing strains of Kluyveromyces lactis have only 43%–55% of the wild-type level of ribosomal DNA (rDNA) repeats. When subjected to prolonged vegetative growth these strains can increase both the number of rDNA repeats and their growth rate.
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    URL: http://dx.doi.org/10.1007/BF00265074
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    Identification of CBS2 as a mitochondrial protein in Saccharomyces cerevisiae (1990)
    Springer
    Molecular genetics and genomics 223 (1990), S. 394-400 
    Details
    ISSN: 1617-4623
    Keywords: Yeast ; Mitochondria ; CBS2 antibodies ; CBS2 protein ; In vitro import
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary The nuclear genome encoded yeast protein CBS2 is required for translational activation of mitochondrial cytochrome b RNA. Genetic studies have shown that the target sequence of the CBS2 protein is the 5′ untranslated leader sequence of cytochrome b RNA. Here we report on the intracellular localization of CBS2. CBS2 protein, expressed in Escherichia coli and prepared from inclusion bodies, was used as an antigen to raise a polyclonal rabbit antiserum. Affinity-purified CBS2 antibodies detect a 45 kDa protein in mitochondrial lysates of wild-type cells, which is absent in a strain in which the CBS2 gene has been deleted. The protein is overexpressed in mitochondrial extracts of a transformant carrying the CBS2 gene on a high copy number plasmid, but undetectable in the post-mitochondrial supernatant. Intramitochondrial localization of CBS2 was verified by in vitro import of CBS2 protein that had been synthesized in a reticulocyte lysate programmed with CBS2 mRNA transcribed in vitro. Mitochondrial import of CBS2 is not accompanied by any detectable proteolytic processing.
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    URL: http://dx.doi.org/10.1007/BF00264445
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    Deletion analysis of the ARG4 promoter of Saccharomyces cerevisine: A poly(dAdT) stretch involved in gene transcription (1990)
    Springer
    Molecular genetics and genomics 223 (1990), S. 474-480 
    Details
    ISSN: 1617-4623
    Keywords: Yeast ; Promoter ; Argininosuccinate lyase ; ARG4
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Transcription of the ARG4 gene of Saccharomyces cerevisiae is regulated by general control of amino acid biosynthesis but not by a specific regulatory mechanism. Three deletion mutants (ΔI, ΔII,, ΔIII) successively removing DNA sequences upstream from the coding sequence have been phenotypically analyzed after insertion into a single copy plasmid. As expected, ΔI, which lacks the sequences upstream to −155, including the two putative upstream activation sequences (UAS), was unable to derepress argininosuccinate lyase biosynthesis under conditions of amino acid starvation. In ΔII (deleted up to −126) the enzyme activity was very low and cells harbouring this allele were arginine dependent. These drastic phenotypic changes can be attributed to the loss of 12 out of 14 dA residues from positions −124 to −137. This poly (dAdT) sequence most likely serves as an upstream promoter element for constitutive expression of ARG4. The ΔIII deletion removes all 5′ sequences including the putative TATA box. This inactive allele has been successfully used for selecting yeast promoters of unknown origin.
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    URL: http://dx.doi.org/10.1007/BF00264456
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    Repair of alkylation damage in Saccharomyces cerevisiae (1990)
    Springer
    Molecular genetics and genomics 221 (1990), S. 353-357 
    Details
    ISSN: 1617-4623
    Keywords: Yeast ; DNA alkylation ; DNA repair
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Repair of methylated bases in Saccharomyces cerevisiae was measured by two methods: in vitro in cell extracts, and in vivo, by determining the loss of methylated bases from yeast DNA after treatment of stationary cultures with [3H]-N-methyl-N′-nitro-N-nitrosoguanidine. Whereas no repair activity could be detected by the in vitro method, the methylated bases were removed in vivo very efficiently. These contradictory results of in vitro and in vivo repair measurements suggest that either the repair enzymes of yeast are sufficiently different from those of bacteria and mammalian cells that they are not active in the in vitro assay, or that methylated bases are repaired in yeast by a different pathway.
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    URL: http://dx.doi.org/10.1007/BF00259399
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  • 35
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    Transcription of two divergently transcribed yeast genes initiates at a common oligo(dA-dT) tract (1990)
    Springer
    Molecular genetics and genomics 223 (1990), S. 438-442 
    Details
    ISSN: 1617-4623
    Keywords: Transcription ; Promoter ; Oligo(dA-dT) stretch ; Gel shift assays ; Yeast
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Oligo(dA-dT) tracts are frequently found in the intergenic regions of the yeast Saccharomyces cerevisiae and have been proposed to act as upstream promoter elements for constitutive transcription. An oligo(dA-dT) tract of 23 bp is also found as a characteristic sequence motif in the centre of the 230 by segment which separates the open reading frames of the CBS2 gene and its 5′-flanking gene on chromosome IV. Recently we have reported that transcription of CBS2 is initiated immediately adjacent to this oligo(dA-dT) tract (Michaelis et al. 1988). Here we report that the flanking gene of unknown function is divergently transcribed into an RNA with heterogeneous 5′ ends. Two of these 5′ ends map within the oligo(dA-dT) stretch, while the third is located upstream, leading to an RNA species which is partially complementary to the CBS2 transcript. Gel shift assays show that the oligo(dA-dT) stretch is specifically recognized by (a) binding factor(s) in nuclear extracts. We discuss these results with respect to the role of oligo(dA-dT) stretches in gene expression in yeast.
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    URL: http://dx.doi.org/10.1007/BF00264451
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