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  • 1
    Electronic Resource
    Electronic Resource
    Chs1 of Candida albicans is an essential chitin synthase required for synthesis of the septum and for cell integrity (2001)
    Oxford, UK : Blackwell Science, Ltd
    Molecular microbiology 39 (2001), S. 0 
    Details
    ISSN: 1365-2958
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology , Medicine
    Notes: CaCHS1 of the fungal pathogen Candida albicans encodes an essential chitin synthase that is required for septum formation, viability, cell shape and integrity. The CaCHS1 gene was inactivated by first disrupting one allele using the ura-blaster protocol, then placing the remaining allele under the control of the maltose-inducible, glucose-repressible MRP1 promoter. Under repressing conditions, yeast cell growth continued temporarily, but daughter buds failed to detach from parents, resulting in septumless chains of cells with constrictions defining contiguous compartments. After several generations, a proportion of the distal compartments lysed. The conditional Δchs1 mutant also failed to form primary septa in hyphae; after several generations, growth stopped, and hyphae developed swollen balloon-like features or lysed at one of a number of sites including the hyphal apex and other locations that would not normally be associated with septum formation. CHS1 therefore synthesizes the septum of both yeast and hyphae and also maintains the integrity of the lateral cell wall. The conditional mutant was avirulent under repressing conditions in an experimental model of systemic infection. Because this gene is essential in vitro and in vivo and is not present in humans, it represents an attractive target for the development of antifungal compounds.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1046/j.1365-2958.2001.02347.x
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  • 2
    Electronic Resource
    Electronic Resource
    Differential post-transcriptional regulation of yeast mRNAs in response to high and low glucose concentrations (2000)
    Oxford, UK : Blackwell Science Ltd
    Molecular microbiology 35 (2000), S. 0 
    Details
    ISSN: 1365-2958
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology , Medicine
    Notes: Glucose regulates yeast gene expression at both transcriptional and post-transcriptional levels. Glucose strongly represses the transcription of the gluconeogenic genes, FBP1 and PCK1, and accelerates the degradation of their mRNAs. Together these mechanisms are responsible for the rapid decrease in gluconeogenic enzyme synthesis when yeast cells switch to glycolytic metabolism. In this study, we show that accelerated gluconeogenic mRNA degradation can be triggered by low concentrations of glucose (〈0.02%). This sets the FBP1 and PCK1 mRNAs apart from other glucose-sensitive mRNAs, such as the Ip mRNA, which only responds to high glucose concentrations (〉1%). We also show that accelerated gluconeogenic mRNA degradation is co-ordinated with transcriptional repression by common signalling components that include sugar kinases and Ras-cAMP signalling. Furthermore, the ability of the low glucose signal to trigger accelerated gluconeogenic mRNA degradation depends upon the low glucose sensor, Snf3p, but not on the high glucose sensor, Rgt2p. Also, this response is influenced by reg1 and ume5 mutations, but not by grr1 or rgt1 mutations. Our data suggest that several signalling pathways co-ordinate differential post-transcriptional and transcriptional responses in yeast, depending upon the amount of glucose available in the medium.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1046/j.1365-2958.2000.01723.x
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  • 3
    Electronic Resource
    Electronic Resource
    Multiple signalling pathways trigger the exquisite sensitivity of yeast gluconeogenic mRNAs to glucose (1996)
    Oxford, UK : Blackwell Publishing Ltd
    Molecular microbiology 20 (1996), S. 0 
    Details
    ISSN: 1365-2958
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology , Medicine
    Notes: The transcription of the yeast FBP1 and PCK1 genes, which encode the gluconeogenic enzymes fructose-1,6-bisphosphatase and phosphoenolpyruvate car-boxykinase, is repressed by glucose. Here, we show that this repression is both very strong and exceptionally sensitive to glucose, being triggered by glucose at concentrations less than 0.005% (0.27 mM). This repression remains operative in yeast mutants carrying any one of the three hexose kinases, but is lost in a triple hxk1, hxk2, glk1 mutant. In addition, 2-deoxyglucose can trigger the repression, but 6-deoxy-glucose cannot, suggesting that internalization and phosphorylation of the glucose is essential for repression to occur. While gluconeogenic gene transcription is subject to the Mig1p-dependent pathway of glucose repression, the exquisite response to glucose is maintained in hxk2 and mig1 mutants, suggesting that this pathway is not essential for the response. The response can also be triggered by the addition of exogenous cAMP, suggesting that the Ras/cAMP pathway can mediate repression of the FPB1 and PCK1 mRNAs. However, the response is not dependent upon this pathway because it remains intact in Ras, adenyl cyclase and protein kinase A mutants. The data show that yeast cells can detect very low glucose concentrations in the environment, and suggest that several distinct signalling pathways operate to repress FPB1 and PCK1 transcription in the presence of glucose.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1111/j.1365-2958.1996.tb02514.x
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  • 4
    Electronic Resource
    Electronic Resource
    The Candida albicans CaACE2 gene affects morphogenesis, adherence and virulence (2004)
    Oxford, UK : Blackwell Science Ltd
    Molecular microbiology 53 (2004), S. 0 
    Details
    ISSN: 1365-2958
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology , Medicine
    Notes: Morphogenesis between yeast and hyphal growth is a characteristic associated with virulence in Candida albicans and involves changes in the cell wall. In Saccharomyces cerevisiae, the transcription factor pair Ace2p and Swi5p are key regulators of cell wall metabolism. Here, we have characterized the CaACE2 gene, which encodes the only C. albicans homologue of S. cerevisiae ACE2 and SWI5. Deleting CaACE2 results in a defect in cell separation, increased invasion of solid agar medium and inappropriate pseudohyphal growth, even in the absence of external inducers. The mutant cells have reduced adherence to plastic surfaces and generate biofilms with distinctly different morphology from wild-type cells. They are also avirulent in a mouse model. Deleting CaACE2 has no effect on expression of the chitinase gene CHT2, but expression of CHT3 and the putative cell wall genes CaDSE1 and CaSCW11 is reduced in both yeast and hyphal forms. The CaAce2 protein is localized to the daughter nucleus of large budded cells at the end of mitosis. C. albicans Ace2p therefore plays a major role in morphogenesis and adherence and resembles S. cerevisiae Ace2p in function.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1111/j.1365-2958.2004.04185.x
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  • 5
    Electronic Resource
    Electronic Resource
    Glucose triggers different global responses in yeast, depending on the strength of the signal, and transiently stabilizes ribosomal protein mRNAs (2003)
    Oxford, UK : Blackwell Publishing Ltd
    Molecular microbiology 48 (2003), S. 0 
    Details
    ISSN: 1365-2958
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology , Medicine
    Notes: Glucose exerts profound effects upon yeast physiology. In general, the effects of high glucose concentrations (〉1%) upon Saccharomyces cerevisiae have been studied. In this paper, we have characterized the global responses of yeast cells to very low (0.01%), low (0.1%) and high glucose signals (1.0%) by transcript profiling. We show that yeast is more sensitive to very low glucose signals than was previously thought, and that yeast displays different responses to these different glucose signals. Genes involved in central metabolic pathways respond rapidly to very low glucose signals, whereas genes involved in the biogenesis of cytoplasmic ribosomes generally respond only to glucose concentrations of〉 0.1%. We also show that cytoplasmic ribosomal protein mRNAs are transiently stabilized by glucose, indicating that both transcriptional and post-transcriptional mechanisms combine to accelerate the accumulation of ribosomal protein mRNAs. Presumably, this facilitates rapid ribosome biogenesis after exposure to glucose. However, our data indicate that yeast activates ribosome biogenesis only when sufficient glucose is available to make this metabolic investment worthwhile. In contrast, the regulation of metabolic functions in response to very low glucose signals presumably ensures that yeast can exploit even minute amounts of this preferred nutrient.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1046/j.1365-2958.2003.03478.x
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  • 6
    Electronic Resource
    Electronic Resource
    Positive regulation of the LPD1 gene of Saccharomyces cerevisine by the HAP2/HAP3/HAP4 activation system (1992)
    Springer
    Molecular genetics and genomics 231 (1992), S. 296-303 
    Details
    ISSN: 1617-4623
    Keywords: Saccharomyces cerevisiae ; Lipoamide dehydrogenase ; HAP activation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary The LPD1 gene of Saccharomyces cerevisiae, encoding lipoamide dehydrogenase (LPDH), is subject to catabolite repression. The promoter of this gene contains a number of motifs for DNA-binding transcriptional activators, including three which show strong sequence homology to the core HAP2/HAP3/HAP4 binding motif. Here we report that transcription of LPD1 requires HAP2, HAP3 and HAP4 for release from glucose repression. In the wild-type strain, specific activity of LPDH was increased 12-fold by growth on lactate, 10-fold on glycerol and four- to five-fold on galactose or raffinose, compared to growth on glucose. In hap2, hap3 and hap4 null mutants, the specific activities of LPDH in cultures grown on galactose and raffinose showed only slight induction above the basal level on glucose medium. Similar results were obtained upon assaying for β-galactosidase production in wild-type, or hap2, hap3 or hap4 mutant strains carrying a single copy of the LPD1 promoter fused in frame to the lacZ gene of Escherichia coli and integrated at the URA3 locus. Transcript analysis in wild-type and hap2 mutants confirmed that the HAP2 protein regulates LPD1 expression at the level of transcription in the same way as it does for the CYC1 gene. Site-directed mutagenesis of the putative HAP2/HAP3/HAP4 binding site at −204 relative to the ATG start codon showed that this element was required for full derepression of the LPD1 gene on non-fermetable substrates.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00279803
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  • 7
    Electronic Resource
    Electronic Resource
    Constitutive activation of the Saccharomyces cerevislae mating response pathway by a MAP kinase kinase from Candida albicans (1995)
    Springer
    Molecular genetics and genomics 249 (1995), S. 609-621 
    Details
    ISSN: 1617-4623
    Keywords: MAP kinase kinase ; Ste7 ; Saccharomyces cerevisiae ; Candida albicans
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The HST7 gene of Candida albicans encodes a protein with structural similarity to MAP kinase kinases. Expression of this gene in Saccharomyces cerevisiae complements disruption of the Ste7 MAP kinase kinase required for both mating in haploid cells and pseudohyphal growth in diploids. However, Hst7 expression does not complement loss of either the Pbs2 (Hog4) MAP kinase kinase required for response to high osmolarity, or loss of the Mkk1 and Mkk2 MAP kinase kinases required for proper cell wall biosynthesis. Intriguingly, HST7 acts as a hyperactive allele of STE7; expression of Hst7 activates the mating pathway even in the absence of upstream signaling components including the Ste7 regulator Ste11, elevates the basal level of the pheromone-inducible FUS1 gene, and amplifies the pseudohyphal growth response in diploid cells. Thus Hst7 appears to be at least partially independent of upstream activators or regulators, but selective in its activity on downstream target MAP kinases. Creation of Hst7/Ste7 hybrid proteins revealed that the C-terminal two-thirds of Hst7, which contains the protein kinase domain, is sufficient to confer this partial independence of upstream activators.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00418030
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  • 8
    Electronic Resource
    Electronic Resource
    5′-Secondary structure formation, in constrast to a short string of non-preferred codons, inhibits the translation of the pyruvate kinase mRNA in yeast (1989)
    New York, NY [u.a.] : Wiley-Blackwell
    Yeast 5 (1989), S. 187-198 
    Details
    ISSN: 0749-503X
    Keywords: Yeast ; messenger RNA ; translation ; codon bias ; RNA secondary-structure ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: The effects of poor codon bias and secondary structure formation upon the translation of the pyruvate kinase (PYK1) mRNA have been investigated in Saccharomyces cerevisiae. Following insertion mutagenesis at the 5′-end of the PYK1 coding region, the gene was transformed into yeast, and translation assessed directly in vivo by determining the distribution of the modified PYK1 mRNAs across polysomes fractionated by sucrose density gradient centrifugation. The chromosomally-encoded (wild-type) PYK1 mRNA, and the actin, ribosomal protein L3 and glyceraldehyde-3-phosphate dehydrogenase mRNAs were used to control for minor differences between polysome preparations. An insertion containing 13 non-preferred codons at the 5′-end of the coding region was found to have no significant effect upon PYK1 mRNA translation. In contrast, translation was inhibited by an insertion which increased the formation of secondary structures at the 5′-end of the mRNA (overall ΔG = -36·6 kcal/mol). Control insertions were also analysed to exclude the possibility that alterations to the amino acid sequence of pyruvate kinase affect the translation of its mRNA. These insertions, which introduced preferred codons or restored wild-type levels of secondary structure formation, did not significantly influence PYK1 mRNA translation.
    Additional Material: 6 Ill.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/yea.320050308
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  • 9
    Electronic Resource
    Electronic Resource
    Messenger RNA stability in yeast (1989)
    New York, NY [u.a.] : Wiley-Blackwell
    Yeast 5 (1989), S. 239-257 
    Details
    ISSN: 0749-503X
    Keywords: Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Additional Material: 7 Ill.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/yea.320050405
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  • 10
    Electronic Resource
    Electronic Resource
    The complete sequence of a 7·5 kb region of chromosome III from Saccharomyces cerevisiae that lies between CRY1 and MAT (1991)
    New York, NY [u.a.] : Wiley-Blackwell
    Yeast 7 (1991), S. 761-772 
    Details
    ISSN: 0749-503X
    Keywords: Chromosome III ; strain polymorphisms ; DNA replication ; mRNA stability ; ribosomal proteins ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: We report the sequence of a 7·5 kb region lying between the CRY1 and MAT loci of chromosome III from Saccharomyces cerevisiae. This region lies in the overlap between two major contigs used for the generation of the complete nucleotide sequence of this chromosome. Comparison of this sequence with those reported previously for this overlap [Thierry et al. (1990) Yeast 6, 521; Jia et al. (1991) Yeast 7, 413] reveals 38 nucleotide differences, 45% of which generate changes in the amino acid sequences of the four genes in this region (YCR591, YCR592, YCR521 and YCR522). These differences appear to reflect true sequence polymorphisms between the two yeast strains used to generate the clones used in the sequencing project. Three of the four genes in this region display weak homologies to proteins in the PIR database. Some properties of YCR521 are analogous to those of ribosomal protein genes. However, the functions of all four genes remain obscure.
    Additional Material: 5 Ill.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/yea.320070711
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