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Biochem J. Sep 1, 2003; 374(Pt 2): 513–519.
PMCID: PMC1223596

Protein S-thiolation targets glycolysis and protein synthesis in response to oxidative stress in the yeast Saccharomyces cerevisiae.


The irreversible oxidation of cysteine residues can be prevented by protein S-thiolation, a process by which protein SH groups form mixed disulphides with low-molecular-mass thiols such as glutathione. We report here the target proteins which are modified in yeast cells in response to H(2)O(2). In particular, a range of glycolytic and related enzymes (Tdh3, Eno2, Adh1, Tpi1, Ald6 and Fba1), as well as translation factors (Tef2, Tef5, Nip1 and Rps5) are identified. The oxidative stress conditions used to induce S-thiolation are shown to inhibit GAPDH (glyceraldehyde-3-phosphate dehydrogenase), enolase and alcohol dehydrogenase activities, whereas they have no effect on aldolase, triose phosphate isomerase or aldehyde dehydrogenase activities. The inhibition of GAPDH, enolase and alcohol dehydrogenase is readily reversible once the oxidant is removed. In addition, we show that peroxide stress has little or no effect on glucose-6-phosphate dehydrogenase or 6-phosphogluconate dehydrogenase, the enzymes that catalyse NADPH production via the pentose phosphate pathway. Thus the inhibition of glycolytic flux is proposed to result in glucose equivalents entering the pentose phosphate pathway for the generation of NADPH. Radiolabelling is used to confirm that peroxide stress results in a rapid and reversible inhibition of protein synthesis. Furthermore, we show that glycolytic enzyme activities and protein synthesis are irreversibly inhibited in a mutant that lacks glutathione, and hence cannot modify proteins by S-thiolation. In summary, protein S-thiolation appears to serve an adaptive function during exposure to an oxidative stress by reprogramming metabolism and protecting protein synthesis against irreversible oxidation.

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Selected References

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  • Carmel-Harel O, Storz G. Roles of the glutathione- and thioredoxin-dependent reduction systems in the Escherichia coli and saccharomyces cerevisiae responses to oxidative stress. Annu Rev Microbiol. 2000;54:439–461. [PubMed]
  • Grant CM. Role of the glutathione/glutaredoxin and thioredoxin systems in yeast growth and response to stress conditions. Mol Microbiol. 2001 Feb;39(3):533–541. [PubMed]
  • Rietsch A, Beckwith J. The genetics of disulfide bond metabolism. Annu Rev Genet. 1998;32:163–184. [PubMed]
  • Coan C, Ji JY, Hideg K, Mehlhorn RJ. Protein sulfhydryls are protected from irreversible oxidation by conversion to mixed disulfides. Arch Biochem Biophys. 1992 Jun;295(2):369–378. [PubMed]
  • Thomas JA, Poland B, Honzatko R. Protein sulfhydryls and their role in the antioxidant function of protein S-thiolation. Arch Biochem Biophys. 1995 May 10;319(1):1–9. [PubMed]
  • Klatt P, Lamas S. Regulation of protein function by S-glutathiolation in response to oxidative and nitrosative stress. Eur J Biochem. 2000 Aug;267(16):4928–4944. [PubMed]
  • Grant CM, Perrone G, Dawes IW. Glutathione and catalase provide overlapping defenses for protection against hydrogen peroxide in the yeast Saccharomyces cerevisiae. Biochem Biophys Res Commun. 1998 Dec 30;253(3):893–898. [PubMed]
  • Grant CM, Quinn KA, Dawes IW. Differential protein S-thiolation of glyceraldehyde-3-phosphate dehydrogenase isoenzymes influences sensitivity to oxidative stress. Mol Cell Biol. 1999 Apr;19(4):2650–2656. [PMC free article] [PubMed]
  • Shenton Daniel, Perrone Gabriel, Quinn Kathryn A, Dawes Ian W, Grant Chris M. Regulation of protein S-thiolation by glutaredoxin 5 in the yeast Saccharomyces cerevisiae. J Biol Chem. 2002 May 10;277(19):16853–16859. [PubMed]
  • Davis DA, Newcomb FM, Starke DW, Ott DE, Mieyal JJ, Yarchoan R. Thioltransferase (glutaredoxin) is detected within HIV-1 and can regulate the activity of glutathionylated HIV-1 protease in vitro. J Biol Chem. 1997 Oct 10;272(41):25935–25940. [PubMed]
  • Obin M, Shang F, Gong X, Handelman G, Blumberg J, Taylor A. Redox regulation of ubiquitin-conjugating enzymes: mechanistic insights using the thiol-specific oxidant diamide. FASEB J. 1998 May;12(7):561–569. [PubMed]
  • Klatt P, Molina EP, De Lacoba MG, Padilla CA, Martinez-Galesteo E, Barcena JA, Lamas S. Redox regulation of c-Jun DNA binding by reversible S-glutathiolation. FASEB J. 1999 Sep;13(12):1481–1490. [PubMed]
  • Jung CH, Thomas JA. S-glutathiolated hepatocyte proteins and insulin disulfides as substrates for reduction by glutaredoxin, thioredoxin, protein disulfide isomerase, and glutathione. Arch Biochem Biophys. 1996 Nov 1;335(1):61–72. [PubMed]
  • Chrestensen CA, Starke DW, Mieyal JJ. Acute cadmium exposure inactivates thioltransferase (Glutaredoxin), inhibits intracellular reduction of protein-glutathionyl-mixed disulfides, and initiates apoptosis. J Biol Chem. 2000 Aug 25;275(34):26556–26565. [PubMed]
  • Luikenhuis S, Perrone G, Dawes IW, Grant CM. The yeast Saccharomyces cerevisiae contains two glutaredoxin genes that are required for protection against reactive oxygen species. Mol Biol Cell. 1998 May;9(5):1081–1091. [PMC free article] [PubMed]
  • Garrido Ester Ocón, Grant Chris M. Role of thioredoxins in the response of Saccharomyces cerevisiae to oxidative stress induced by hydroperoxides. Mol Microbiol. 2002 Feb;43(4):993–1003. [PubMed]
  • Rodríguez-Manzaneque MT, Ros J, Cabiscol E, Sorribas A, Herrero E. Grx5 glutaredoxin plays a central role in protection against protein oxidative damage in Saccharomyces cerevisiae. Mol Cell Biol. 1999 Dec;19(12):8180–8190. [PMC free article] [PubMed]
  • Bushweller JH, Aslund F, Wüthrich K, Holmgren A. Structural and functional characterization of the mutant Escherichia coli glutaredoxin (C14----S) and its mixed disulfide with glutathione. Biochemistry. 1992 Sep 29;31(38):9288–9293. [PubMed]
  • Eaton Philip, Byers Helen L, Leeds Nicola, Ward Malcolm A, Shattock Michael J. Detection, quantitation, purification, and identification of cardiac proteins S-thiolated during ischemia and reperfusion. J Biol Chem. 2002 Mar 22;277(12):9806–9811. [PubMed]
  • Fratelli Maddalena, Demol Hans, Puype Magda, Casagrande Simona, Eberini Ivano, Salmona Mario, Bonetto Valentina, Mengozzi Manuela, Duffieux Francis, Miclet Emeric, et al. Identification by redox proteomics of glutathionylated proteins in oxidatively stressed human T lymphocytes. Proc Natl Acad Sci U S A. 2002 Mar 19;99(6):3505–3510. [PMC free article] [PubMed]
  • Lind Christina, Gerdes Robert, Hamnell Ylva, Schuppe-Koistinen Ina, von Löwenhielm Helena Brockenhuus, Holmgren Arne, Cotgreave Ian A. Identification of S-glutathionylated cellular proteins during oxidative stress and constitutive metabolism by affinity purification and proteomic analysis. Arch Biochem Biophys. 2002 Oct 15;406(2):229–240. [PubMed]
  • Grant CM, MacIver FH, Dawes IW. Glutathione is an essential metabolite required for resistance to oxidative stress in the yeast Saccharomyces cerevisiae. Curr Genet. 1996 May;29(6):511–515. [PubMed]
  • McAlister L, Holland MJ. Isolation and characterization of yeast strains carrying mutations in the glyceraldehyde-3-phosphate dehydrogenase genes. J Biol Chem. 1985 Dec 5;260(28):15013–15018. [PubMed]
  • Maitra PK, Lobo Z. A kinetic study of glycolytic enzyme synthesis in yeast. J Biol Chem. 1971 Jan 25;246(2):475–488. [PubMed]
  • Poyner RR, Laughlin LT, Sowa GA, Reed GH. Toward identification of acid/base catalysts in the active site of enolase: comparison of the properties of K345A, E168Q, and E211Q variants. Biochemistry. 1996 Feb 6;35(5):1692–1699. [PubMed]
  • Rippa M, Signorini M. 6-Phosphogluconate dehydrogenase from Candida utilis. Methods Enzymol. 1975;41:237–240. [PubMed]
  • Izawa S, Inoue Y, Kimura A. Oxidative stress response in yeast: effect of glutathione on adaptation to hydrogen peroxide stress in Saccharomyces cerevisiae. FEBS Lett. 1995 Jul 10;368(1):73–76. [PubMed]
  • Stephen DW, Jamieson DJ. Glutathione is an important antioxidant molecule in the yeast Saccharomyces cerevisiae. FEMS Microbiol Lett. 1996 Aug 1;141(2-3):207–212. [PubMed]
  • Godon C, Lagniel G, Lee J, Buhler JM, Kieffer S, Perrot M, Boucherie H, Toledano MB, Labarre J. The H2O2 stimulon in Saccharomyces cerevisiae. J Biol Chem. 1998 Aug 28;273(35):22480–22489. [PubMed]
  • Werner-Washburne M, Braun EL, Crawford ME, Peck VM. Stationary phase in Saccharomyces cerevisiae. Mol Microbiol. 1996 Mar;19(6):1159–1166. [PubMed]
  • Werner-Washburne M, Braun E, Johnston GC, Singer RA. Stationary phase in the yeast Saccharomyces cerevisiae. Microbiol Rev. 1993 Jun;57(2):383–401. [PMC free article] [PubMed]
  • Schuppe-Koistinen I, Moldéus P, Bergman T, Cotgreave IA. S-thiolation of human endothelial cell glyceraldehyde-3-phosphate dehydrogenase after hydrogen peroxide treatment. Eur J Biochem. 1994 May 1;221(3):1033–1037. [PubMed]
  • Ravichandran V, Seres T, Moriguchi T, Thomas JA, Johnston RB., Jr S-thiolation of glyceraldehyde-3-phosphate dehydrogenase induced by the phagocytosis-associated respiratory burst in blood monocytes. J Biol Chem. 1994 Oct 7;269(40):25010–25015. [PubMed]
  • Holmgren A. Thioredoxin and glutaredoxin systems. J Biol Chem. 1989 Aug 25;264(24):13963–13966. [PubMed]
  • Kletzien RF, Harris PK, Foellmi LA. Glucose-6-phosphate dehydrogenase: a "housekeeping" enzyme subject to tissue-specific regulation by hormones, nutrients, and oxidant stress. FASEB J. 1994 Feb;8(2):174–181. [PubMed]
  • Izawa S, Maeda K, Miki T, Mano J, Inoue Y, Kimura A. Importance of glucose-6-phosphate dehydrogenase in the adaptive response to hydrogen peroxide in Saccharomyces cerevisiae. Biochem J. 1998 Mar 1;330(Pt 2):811–817. [PMC free article] [PubMed]
  • Flattery-O'Brien JA, Dawes IW. Hydrogen peroxide causes RAD9-dependent cell cycle arrest in G2 in Saccharomyces cerevisiae whereas menadione causes G1 arrest independent of RAD9 function. J Biol Chem. 1998 Apr 10;273(15):8564–8571. [PubMed]

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