A single glutaredoxin or thioredoxin gene is essential for viability in the yeast Saccharomyces cerevisiae.
Glutaredoxins and thioredoxins are small heat-stable oxidoreductases that have been conserved throughout evolution. The yeast Saccharomyces cerevisiae contains two gene pairs encoding cytoplasmic glutaredoxins (GRX1, GRX2) and thioredoxins (TRX1, TRX2). We report here that the quadruple trx1 trx2 grx1 grx2 mutant is inviable and that either a single glutaredoxin or a single thioredoxin (i.e. grx1 grx2 trx1, grx1 grx2 trx2, grx1 trx1 trx2, grx2 trx1 trx2) is essential for viability. Loss of both thioredoxins has been reported previously to lead to methionine auxotrophy consistent with thioredoxins being the sole reductants for 3'-phosphoadenosine 5'-phosphosulphate reductase (PAPS) in yeast. However, we present evidence for the existence of a novel yeast hydrogen donor for PAPS reductase, as strains lacking both thioredoxins assimilated sulphate under conditions that minimized the generation of reactive oxygen species (low aeration and absence of functional mitochondria). In addition, the assimilation of [35S]-sulphate was approximately 60-fold higher in the trx1 trx2 grx1 and trx1 trx2 grx2 mutants compared with the trx1 trx2 mutant. Furthermore, in contrast to the trx1 trx2 mutant, the trx1 trx2 grx2 mutant grew on minimal agar plates, and the trx1 trx2 grx1 mutant grew on minimal agar plates under anaerobic conditions. We propose a model in which the novel reductase activity normally functions in the repair of oxidant-mediated protein damage but, under conditions that minimize the generation of reactive oxygen species, it can serve as a hydrogen donor for PAPS reductase.
Draculic
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School of Biochemistry and Molecular Genetics, University of New South Wales., Sydney, NSW 2052, Australia.
Dawes
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IW
Grant
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CM
D003470:Culture Media; D018628:Gene Dosage; D005800:Genes, Fungal; D054477:Glutaredoxins; D008565:Membrane Proteins / Q000235:genetics*; D016296:Mutagenesis; D010088:Oxidoreductases / Q000235:genetics*; D054464:Peroxiredoxins; D010641:Phenotype; D011506:Proteins / Q000235:genetics*; D012441:Saccharomyces cerevisiae / Q000201:enzymology* / Q000235:genetics / Q000254:growth & development*; D029701:Saccharomyces cerevisiae Proteins; D013431:Sulfates / Q000378:metabolism; D013879:Thioredoxins / Q000235:genetics*
D003470:Culture Media; D054477:Glutaredoxins; D008565:Membrane Proteins; D011506:Proteins; D029701:Saccharomyces cerevisiae Proteins; D013431:Sulfates; C103243:TRX1 protein, S cerevisiae; C516110:TRX2 protein, S cerevisiae; D013879:Thioredoxins; D010088:Oxidoreductases; D054464:Peroxiredoxins