Oxidative Stresses and cancer
Michel B. TOLEDANO
CEA Saclay/Bât. 142
Tél: 01 69 08 82 44
michel.toledano@cea.fr
CEA Saclay/Bât. 142
Tél: 01 69 08 82 44
michel.toledano@cea.fr
Understand and describe the cellular metabolism of oxygen and reactive oxygen species (ROS) acting on cysteine residues
Human resources
Michel B. TOLEDANO, Group Leader
Agnès DELAUNAY-MOISAN, Researcher
Stéphanie LURIAU, Research Technician
Marie-Elyse LANCELOT, Research Technician
Gael PALAIS, Research Technician
Research Programs
We study the biological effects of oxygen and its reactive species, called ROS, including the superoxide ion and hydrogen peroxide, as well as the redox reactions acting on cysteine residues. The deregulation of biological systems affected by oxygen and ROS or implicated in their metabolism is an essential factor in aging and associated diseases, in particular cancer.
Our environment is oxidizing because of the prevalence of ambient oxygen. This environmental characteristic, if not countered, would lead ineluctably to the oxidation of all organic matter on earth. Aerobic organisms have domesticated the oxidizing power of O2 in utilizing it as the terminal electron acceptor of the respiratory chain generating cellular energy, and as a catalyst or cofactor of a multitude of reactions utilizing the redox properties of cysteine residues in many cases. Examples include DNA synthesis (ribonucleotide reductase), the folding of secreted proteins by disulfide bond formation catalyzed by the Ero1 oxydase, the import of certain mitochondrial proteins by the formation of disulfide bonds catalyzed by Erv1, cell signaling by H2O2, etc. The domestication of oxygen is equally linked to cellular ROS detoxification systems that use cysteine residues to reduce H2O2, such as the peroxiredoxines (Prxs) and glutathione peroxidases (GPxs). Cysteine is used in these reactions because it is found alternatively in two redox forms, the reduced thiol (-SH) and the oxidized disulfide bond (S-S). An essential theme of our group is to study the ability of cysteine oxidation to act as a regulatory mechanism through its potential to impact protein conformation. Two cellular systems control the oxidation state of cysteine, the glutathione and thioredoxin pathways, and they use the redox properties of cysteine themselves for this function.
Research projects :
1) Signaling pathways controlling anti-oxydant systems. These pathways are activated by H2O2 and they are composed of ultra-precise detection mechanisms for this oxydant (see Fig. 1).
Yap1 in S. cerevisiae. We showed that Yap1 is activated by oxidation. We identified Orp1 as the protein sensor detecting H2O2 and oxidizing Yap1. Our efforts are now concentrated on a structure-function analysis of the Yap1 system in vivo and in vitro after reconstitution with purified proteins.a)
b) The Keap1-Nrf2 pathway in mammals. We would like to understand how Keap1, controlling the degradation of the transcription factor Nrf2, is regulated by H2O2 and by nitric oxide (NO).
2) The identification of proteins containing oxidized cysteine residues and the targets of the GSH and thioredoxin pathways at the level of the total S. cerevisiae proteome. Our objective is to identify novel redox mechanisms.
3) The biological function of anti-oxydants.
a) In S. cerevisiae: establish the function of all the systems contributing to the cellular tolerance to H2O2: the anti-oxidants, the GSH and thioredoxin pathways, and the regulators implicated in the control of this tolerance. This systems biology project benefits from innovative genetic approaches.
b) In mice: establish knockout mice inactivated for genes encoding anti-oxidants and DNA repair enzymes to determine the toxicity of ROS with regards to DNA and carcinogenicity.
©CEA/M.B. ToledanoFigure 1. Model for the mechanism of H2O2 detection by Orp-Yap1. Yap1 forms a pre-complex with the Ybp1 chaperone. Orp1 is oxidized by H2O2 on Cys36. The resulting Cys36-SOH (sulfenic acid) condenses with Cys598 of Yap1 to form an intermolecular disulfide bond that is then transformed into an intramolecular disulfide bond by attack of Yap1-Cys303. Ybp1 acts as a scaffold to facilitate the Orp1-Yap1 redox interaction. The activation of Yap1 by electrophiles and metals is Orp1-independent. The peroxidative cycle of Orp1 is also shown (see Delaunay et al. (2002) for further details).
©CEA/F. Tacnet & M. B. Toledano
Figure 2. Use of an H2O2-specific fluorescent reporter to quantify the intracellular concentration of this oxydant in yeast cells. Note the remarkable heterogeneity of this concentration within cells. Unpublished data of Tacnet and Toledano.
Key words
Signaling and stress detection, redox regulation, oxidative stress, hydrogen peroxide, anti-oxidants, redox homeostasis, disulfide bond, cancer, aging, post-translational modification, Saccharomyces cerevisiae, mammals, transgenic mice, genetic screens, protein biochemistry, cell biology, proteomics, systems biology.
Publications
Boisnard S, Lagniel G, Garmendia-Torres C, Molin M, Boy-Marcotte E, Jacquet M, Toledano MB, Labarre J, Chédin S. (2009). H2O2 activates the nuclear localization of Msn2 and Maf1 through thioredoxins in Saccharomyces cerevisiae. Eukaryot Cell. 8, 1429-38
Kaur H, Kumar C, Junot C, Toledano MB, Bachhawat AK. (2009). Dug1p Is a Cys-Gly peptidase of the gamma-glutamyl cycle of Saccharomyces cerevisiae and represents a novel family of Cys-Gly peptidases. J Biol Chem. 284, 14493-14502.
Fourquet S, Huang ME, D'Autreaux B, Toledano MB. (2008). The Dual Functions of Thiol-Based Peroxidases in H(2)O(2) Scavenging and Signaling. Antioxid Redox Signal. 10, 1565-76.
Guerrier L, D'Autréaux B, Atanassov C, Khoder G, Boschetti E. (2008). Evaluation of a standardized method of protein purification and identification after discovery by mass spectrometry. J Proteomics. 71, 368-78.
Le Moan N, Tacnet F, Toledano MB. (2008). Protein-thiol oxidation, from single proteins to proteome-wide analyses. Methods Mol Biol. 476, 181-98.
Azevedo D, Nascimento L, Labarre J, Toledano MB, Rodrigues-Pousada C. (2007). The S. cerevisiae Yap1 and Yap2 transcription factors share a common cadmium-sensing domain FEBS Lett. 581, 187-195.
Camier S, Ma E, Leroy C, Pruvost A, Toledano M, Marsolier-Kergoat MC. (2007). Visualization of ribonucleotide reductase catalytic oxidation establishes thioredoxins as its major reductants in yeast. Free Radic Biol Med. 42, 1008-10016.
D'Autréaux B, Toledano MB. (2007). ROS as signalling molecules: mechanisms that generate specificity in ROS homeostasis. Nat Rev Mol Cell Biol. 8, 813-24
Diet A, Abbas K, Bouton C, Guillon B, Tomasello F, Fourquet S, Toledano MB, Drapier JC. (2007).Regulation of peroxiredoxins by nitric oxide in immunostimulated macrophages. J Biol Chem. 282, 36199-205.
Lopez-Mirabal HR, Thorsen M, Kielland-Brandt MC, Toledano MB, Winther JR. (2007).Cytoplasmic glutathione redox status determines survival upon exposure to the thiol-oxidant 4,4'-dipyridyl disulfide. FEMS Yeast Res. 7, 391-403.
Kaur H, Kumar C, Junot C, Toledano MB, Bachhawat AK. (2009). Dug1p Is a Cys-Gly peptidase of the gamma-glutamyl cycle of Saccharomyces cerevisiae and represents a novel family of Cys-Gly peptidases. J Biol Chem. 284, 14493-14502.
Fourquet S, Huang ME, D'Autreaux B, Toledano MB. (2008). The Dual Functions of Thiol-Based Peroxidases in H(2)O(2) Scavenging and Signaling. Antioxid Redox Signal. 10, 1565-76.
Guerrier L, D'Autréaux B, Atanassov C, Khoder G, Boschetti E. (2008). Evaluation of a standardized method of protein purification and identification after discovery by mass spectrometry. J Proteomics. 71, 368-78.
Le Moan N, Tacnet F, Toledano MB. (2008). Protein-thiol oxidation, from single proteins to proteome-wide analyses. Methods Mol Biol. 476, 181-98.
Azevedo D, Nascimento L, Labarre J, Toledano MB, Rodrigues-Pousada C. (2007). The S. cerevisiae Yap1 and Yap2 transcription factors share a common cadmium-sensing domain FEBS Lett. 581, 187-195.
Camier S, Ma E, Leroy C, Pruvost A, Toledano M, Marsolier-Kergoat MC. (2007). Visualization of ribonucleotide reductase catalytic oxidation establishes thioredoxins as its major reductants in yeast. Free Radic Biol Med. 42, 1008-10016.
D'Autréaux B, Toledano MB. (2007). ROS as signalling molecules: mechanisms that generate specificity in ROS homeostasis. Nat Rev Mol Cell Biol. 8, 813-24
Diet A, Abbas K, Bouton C, Guillon B, Tomasello F, Fourquet S, Toledano MB, Drapier JC. (2007).Regulation of peroxiredoxins by nitric oxide in immunostimulated macrophages. J Biol Chem. 282, 36199-205.
Lopez-Mirabal HR, Thorsen M, Kielland-Brandt MC, Toledano MB, Winther JR. (2007).Cytoplasmic glutathione redox status determines survival upon exposure to the thiol-oxidant 4,4'-dipyridyl disulfide. FEMS Yeast Res. 7, 391-403.
Molin M, Renault JP, Lagniel G, Pin S, Toledano M, Labarre J. (2007).Ionizing radiation induces a Yap1-dependent peroxide stress response in yeast. Free Radic Biol Med. 43, 136-144.
Nadeau PJ, Charette SJ, Toledano MB, Landry J. (2007). Disulfide Bond-mediated Multimerization of Ask1 and Its Reduction by Thioredoxin-1 Regulate H2O2-induced JNK Activation and Apoptosis. Mol Biol Cell. 18, 3903-13
Rey P, Becuwe N, Barrault MB, Rumeau D, Havaux M, Biteau B, Toledano MB. (2007). The Arabidopsis thaliana sulfiredoxin is a plastidic cysteine-sulfinic acid reductase involved in the photooxidative stress response. Plant J. 49, 505-514.
Toledano, M. B., Kumar, C., Le Moan, N., Spector, D., Tacnet, F. (2007). The system biology of thiol redox system in E. coli and yeast: differential functions in oxidative stress, iron metabolism and DNA synthesis. FEBS Let, 581, 3598-607 (Erratum in: FEBS Lett. 2007 Sep 18;581(23):45).
Le Moan N, Clement G, Le Maout S, Tacnet F, Toledano MB. (2006). The Saccharomyces cerevisiae proteome of oxidized protein thiols: contrasted functions for the thioredoxin and glutathione pathways J Biol Chem. 281, 10420-10430.
Vivancos A, Castillo E, Bîteau B, Nicot C, Ayté J, Toledano M B, Hidalgo E. (2005). A cysteine-sulfinic acid in peroxiredoxin regulates H2O2 sensing by the antioxidant Pap1 pathway. Proc Natl Acad Sci U S A. 102, 8875–8880.
Toledano M, Delaunay A, Monceau L, Tacnet F. (2004). Microbial H2O2 sensors as archetypical redox signaling modules Trends Biochem Sci. 29, 351-357.
Menezes RA, Amaral C, Delaunay A, Toledano M, Rodrigues-Pousad C. (2004). Yap8p activation in Saccharomyces cerevisiae under arsenic conditions. FEBS Lett. 566, 141-146.
Desaint S, Luriau S, Aude JC, Rousselet G, Toledano MB. (2004). Mammalian antioxidant defenses are not inducible by H2O2. J Biol Chem. 279, 31157-31163.
Toledano MB, Delaunay A, Biteau B, Spector D, Azevedo D. (2003). Oxidative stress responses in yeast. Topics Cur Genet. 1, 241-303.
Biteau B, Labarre J, Toledano MB. (2003). ATP-dependent reduction of cysteine-sulphinic acid by S-cerevisiae sulphiredoxin. Nature. 425, 980-984.
Azevedo D, Tacnet F, Delaunay A, Rodrigues-Pousada C, Toledano MB. (2003). Two redox centers within Yap1 for H2O2 and thiol-reactive chemicals signalling. Free Radic Biol Med. 35, 889-900.
Hasan R, Leroy C, Isnard AD, Labarre J, Boy-Marcotte E, Toledano MB.(2002). The control of the yeast H2O2 response by the Msn2/4 transcription factors. Mol Microbiol. 45, 233-245.
Delaunay A, Pflieger D, Barrault MB, Vinh J, Toledano MB.(2002). A thiol peroxidase is an H2O2 receptor and redox-transducer in gene activation. Cell. 111, 471-481.
Vido K, Spector D, Lagniel G, Lopez S, Toledano MB, Labarre J. (2001). A proteome analysis of the cadmium response in Saccharomyces cerevisiae. J Biol Chem. 276, 8469-8474.
Spector D, Labarre J, Toledano MB. (2001). A genetic investigation of the essential role of glutathione. Mutations in the proline biosynthesis pathway are the only suppressors of glutathione auxotrophy in yeast. J Biol Chem. 276, 7011-7016.
Meyrial V, Laize V, Gobin R, Ripoche P, Hohmann S, Tacnet F. (2001). Existence of a tightly regulated water channel in Saccharomyces cerevisiae. Eur J Biochem. 268, 334-343.
Laizé V, Tacnet F, Ripoche P, Hohmann S. (2000). Polymorphism of Saccharomyces cerevisiae aquaporins. Yeast. 16, 897-903.
Delaunay A, Isnard AD, Toledano MB. (2000). H2O2 sensing through oxidation of the Yap1 transcription factor. EMBO J. 19, 5157-5166.
Lee J, Godon C, Lagniel G, Spector D, Garin J, Labarre J, Toledano MB. (1999). Yap1 and Skn7 control two specialized oxidative stress response regulons in yeast. J Biol Chem. 274, 16040-16046
Lee J, Spector D, Godon C, Labarre J, Toledano MB. (1999). A new antioxidant with alkyl hydroperoxide defense properties in yeast.J Biol Chem. 274, 4537-4544.
Laizé V, Gobin R, Rousselet G, Badier C, Hohmann S, Ripoche P, Tacnet F. (1999). Molecular and functional study of AQY1 from Saccharomyces cerevisiae : role of the C-terminal domain. Biochem Biophys Res Commun. 257, 139-144.