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  • 1
    ISSN: 1432-0983
    Keywords: Saccharomyces cerevisiae ; Allosuppressor ; Translation ; Fidelity
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Allosuppressor (sal) mutations enhance the efficiency of the yeast ochre suppressor SUQ5 and define five unlinked loci, SALT-SALS. A number of sal4 mutants were isolated and found to have pleiotropic, allele;specific phenotypes, including hypersensitivity in vivo to paromomycin and other antibiotics that stimulate translational errors in yeast. To examine further the nature of the SAL4 gene product, the wild type SAL4 gene was isolated by complementation of a conditional lethal allele sal4-2, and demonstrated to be a single copy gene encoding a single 1.6 kb transcript. Restriction mapping and DNA hybridisation analysis were used to demonstrate that the SAL4 gene is identical to the previously identified omnipotent suppressor gene SUP45 (SUPT). Our results implicate the SAL4 gene product as playing a major role in maintaining translational accuracy in yeast.
    Type of Medium: Electronic Resource
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  • 2
    ISSN: 1432-0983
    Keywords: Key words  Oxidative stress ; Glutathione ; Yeast
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract   Glutathione (GSH) is an abundant cellular thiol which has been implicated in numerous cellular processes and in protection against stress caused by xenobiotics, carcinogens and radiation. Our experiments address the requirement for GSH in yeast, and its role in protection against oxidative stress. Mutants which are unable to synthesis GSH due to a gene disruption in GSH 1, encoding the enzyme for the first step in the biosynthesis of GSH, require exogenous GSH for growth under non-stress conditions. Growth can also be restored with reducing agents containing a sulphydryl group, including dithiothreitol, β-mercaptoethanol and cysteine, indicating that GSH is essential only as a reductant during normal cellular processes. In addition, the GSH 1-disruption strain is sensitive to oxidative stress caused by H2O2 and tert-butyl hydroperoxide. The requirement for GSH in protection against oxidative stress is analogous to that in higher eukaryotes, but unlike the situation in bacteria where it is dispensable for growth during both normal and oxidative stress conditions.
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  • 3
    ISSN: 1432-0983
    Keywords: Key wordsCOX1 ; Intron ; Yeast ; Promoter
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Screening of a promoter probe gene bank for DNA sequences that could act as inducible promoters following growth on non-fermentable carbon sources led to the identification of the mitochondrially encoded cytochrome oxidase subunit 1 gene (COX1) as an active sequence. Carbon-source regulation of this promoter was confirmed by a β-galactosidase assay which showed a 31- and 180-fold induction of expression after growth on ethanol or lactate-based media respectively. Two elements matching the CCAAT-binding-factor motif, which is involved in activating transcription on non-fermentable carbon sources, were identified in the putative promoter. Expression was found to be reduced to low levels in otherwise isogenic hap3 and hap4 mutant strains. Thus, this mitochondrial DNA when placed in the nucleus can act as a promoter that is subject to strict carbon-source regulation. These observations are discussed both with respect to the origin of the S. cerevisiae COX1 gene in particular and with respect to the origin of introns in general.
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  • 4
    Electronic Resource
    Electronic Resource
    Oxford, UK : Blackwell Publishing Ltd
    Molecular microbiology 3 (1989), S. 0 
    ISSN: 1365-2958
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology , Medicine
    Notes: The synthesis of heat-shock proteins can be triggered by a variety of stress-inducing conditions. Here we show that translational misreading caused by growth in the presence of the aminoglycoside antibiotic paromomycin will induce the heat-shock response in the yeast Saccharomyces cerevisiae. This was demonstrated (i) by the acquisition of thermotolerance, and (ii) by elevated levels of expression of the heat-shock protein, hsp70. In addition, transcription of the ubiquitin gene (UBI4) was increased in paromomycin-grown cells. Control experiments with the protein synthesis inhibitor cycloheximide (which does not induce translational misreading) demonstrated that the response was not due to inhibition of protein synthesis per se. These observations strongly suggest that the synthesis of abnormally high levels of aberrant proteins is the trigger of the heat-shock response in this simple eukaryote.
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