Modulation of the specific glutathionylation of mitochondrial proteins in the yeastSaccharomyces cerevisiaeunder basal and stress conditions.
The potential biological consequences of oxidative stress and changes in glutathione levels include the oxidation of susceptible protein thiols and reversible covalent binding of glutathione to the -SH groups of proteins by S-glutathionylation. Mitochondria are central to the response to oxidative stress and redox signaling. It is therefore crucial to explore the adaptive response to changes in thiol-dependent redox status in these organelles. We optimized the purification protocol of glutathionylated proteins in the yeastSaccharomyces cerevisiaeand present a detailed proteomic analysis of the targets of protein glutathionylation in cells undergoing constitutive metabolism and after exposure to various stress conditions. This work establishes the physiological importance of the glutathionylation process inS. cerevisiaeunder basal conditions and provides evidence for an atypical and unexpected cellular distribution of the process between the cytosol and mitochondria. In addition, our data indicate that each oxidative condition (diamide, GSSG, H2O2, or the presence of iron) elicits an adaptive metabolic response affecting specific mitochondrial metabolic pathways, mainly involved in the energetic maintenance of the cells. The correlation of protein modifications with intracellular glutathione levels suggests that protein deglutathionylation may play a role in protecting mitochondria from oxidative stress. This work provides further insights into the diversity of proteins undergoing glutathionylation and the role of this post-translational modification as a regulatory process in the adaptive response of the cell.
Laboratoire Mitochondries, Métaux et Stress Oxydant, Institut Jacques Monod, UMR 7592, Université Paris-Diderot/CNRS, 15 rue Hélène Brion, 75013 Paris, France.
Rachel
R
Gergondey
Plateforme Protéomique structurale et fonctionnelle/Spectrométrie de masse, Institut Jacques Monod, UMR 7592, Université Paris-Diderot/CNRS, 15 rue Hélène Brion, 75013 Paris, France.
Camille
C
Garcia
Laboratoire de Biologie Moléculaire et Cellulaire des Eucaryotes, Institut de Biologie Physico-Chimique, UMR 8226, CNRS, Sorbonne Universités, UPMC Université Paris 6, 13 rue Pierre et Marie Curie, 75005 Paris, France.
Christophe H
CH
Marchand
Laboratoire de Biologie Moléculaire et Cellulaire des Eucaryotes, Institut de Biologie Physico-Chimique, UMR 8226, CNRS, Sorbonne Universités, UPMC Université Paris 6, 13 rue Pierre et Marie Curie, 75005 Paris, France.
Stephane D
SD
Lemaire
Laboratoire Mitochondries, Métaux et Stress Oxydant, Institut Jacques Monod, UMR 7592, Université Paris-Diderot/CNRS, 15 rue Hélène Brion, 75013 Paris, France.
Jean-Michel
JM
Camadro
Laboratoire Mitochondries, Métaux et Stress Oxydant, Institut Jacques Monod, UMR 7592, Université Paris-Diderot/CNRS, 15 rue Hélène Brion, 75013 Paris, France francoise.auchere@ijm.fr.
Françoise
F
Auchère
D003600:Cytosol / Q000187:drug effects / Q000378:metabolism; D003958:Diamide / Q000494:pharmacology; D063990:Gene Ontology; D005978:Glutathione / Q000378:metabolism*; D019803:Glutathione Disulfide / Q000494:pharmacology; D006861:Hydrogen Peroxide / Q000494:pharmacology; D007501:Iron / Q000378:metabolism; D008928:Mitochondria / Q000187:drug effects / Q000378:metabolism; D024101:Mitochondrial Proteins / Q000235:genetics / Q000378:metabolism*; D058977:Molecular Sequence Annotation; D010084:Oxidation-Reduction; D018384:Oxidative Stress; D011499:Protein Processing, Post-Translational; D040901:Proteomics; D012441:Saccharomyces cerevisiae / Q000187:drug effects / Q000235:genetics / Q000378:metabolism*; D029701:Saccharomyces cerevisiae Proteins / Q000235:genetics / Q000378:metabolism*
D024101:Mitochondrial Proteins; D029701:Saccharomyces cerevisiae Proteins; D003958:Diamide; D006861:Hydrogen Peroxide; D007501:Iron; D005978:Glutathione; D019803:Glutathione Disulfide
Saccharomyces cerevisiae; glutathione; glutathionylation; proteomics; redox status