Hydrogen peroxide-induced carbonylation of key metabolic enzymes in Saccharomyces cerevisiae: the involvement of the oxidative stress response regulators Yap1 and Skn7

• Published in: Free radical biology & medicine 2002-12, Vol.33 (11), p.1507-1515
• Main Authors: Costa, Vítor M.V, Amorim, Maria Amélia, Quintanilha, Alexandre, Moradas-Ferreira, Pedro
• Format: Article
• Language: English
• Subjects: Carbon - metabolism; CuZn-superoxide dismutase; Cyclophilin 1; DNA-Binding Proteins - metabolism; Electrophoresis, Gel, Two-Dimensional; Free radicals; Glyceraldehyde-3-phosphate dehydrogenase; Glyceraldehyde-3-Phosphate Dehydrogenase (Phosphorylating) - metabolism; Glyceraldehyde-3-Phosphate Dehydrogenases - metabolism; Hydrogen peroxide; Hydrogen Peroxide - pharmacology; Oxidative Stress; Oxygen - metabolism; Phosphoglycerate mutase; Phosphoglycerate Mutase - metabolism; Protein carbonylation; Protein Isoforms; Saccharomyces cerevisiae - enzymology; Saccharomyces cerevisiae - metabolism; Saccharomyces cerevisiae Proteins - metabolism; Skn7; Superoxide Dismutase - metabolism; Transcription Factors - metabolism; Triose-Phosphate Isomerase - metabolism; Yap1; Yeast
• ISSN: 0891-5849; 1873-4596
• DOI: 10.1016/S0891-5849(02)01086-9

H 2O 2 induces a specific protein oxidation in yeast cells, and the glycolytic enzyme glyceraldehyde-3-phosphate dehydrogenase (Tdh) is a major target. Using a 2D-gel system to study protein carbonylation, it is shown in this work that both Tdh2p and Tdh3p isozymes were oxidized during exposure to H 2O 2. In addition, we identified two other proteins carbonylated and inactivated: Cu,Zn-superoxide dismutase and phosphoglycerate mutase. The oxidative inactivation of Cu,Zn-superoxide dismutase decreases the antioxidant capacity of yeast cells and probably contributes to H 2O 2-induced cell death. Cyclophilin 1 was also carbonylated, but CPH1 gene disruption did not affect peroxide stress sensitivity. The correlation between H 2O 2 sensitivity and the accumulation of oxidized proteins was evaluated by assaying protein carbonyls in mutants deficient in the stress response regulators Yap1p and Skn7p. The results show that the high sensitivity of yap1Δ and skn7Δ mutants to H 2O 2 was correlated with an increased induction of protein carbonylation. In wild-type cells, the acquisition of stress resistance by pre-exposure to a sublethal H 2O 2 stress was associated with a lower accumulation of oxidized proteins. However, pre-exposure of yap1Δ and skn7Δ cells to 0.4 mM H 2O 2 decreased protein carbonylation induced by 1.5 mM H 2O 2, indicating that the adaptive mechanism involved in the protection of proteins from carbonylation is Yap1p- and Skn7p-independent.