Simon Fourquet

The NRF2 transcription factor regulates a major environmental and oxidative stress response. NRF2 is itself negatively regulated by KEAP1, the adaptor of a Cul3-ubiquitin ligase complex that marks NRF2 for proteasomal degradation by ubiquitination. Electrophilic compounds activate NRF2 primarily by inhibiting KEAP1-dependent NRF2 degradation, through alkylation of specific cysteines. We have examined the impact on KEAP1 of reactive oxygen and nitrogen species, which are also NRF2 inducers. We found that in untreated cells, a fraction of KEAP1 carried a long range disulfide linking Cys(226) and Cys(613). Exposing cells to hydrogen peroxide, to the nitric oxide donor spermine NONOate, to hypochlorous acid, or to S-nitrosocysteine further increased this disulfide and promoted formation of a disulfide linking two KEAP1 molecules via Cys(151). None of these oxidants, except S-nitrocysteine, caused KEAP1 S-nitrosylation. A cysteine mutant preventing KEAP1 intermolecular disulfide formation also prevented NRF2 stabilization in response to oxidants, whereas those preventing intramolecular disulfide formation were functionally silent. Further, simultaneously inactivating the thioredoxin and glutathione pathways led both to major constitutive KEAP1 oxidation and NRF2 stabilization. We propose that KEAP1 intermolecular disulfide formation via Cys(151) underlies the activation of NRF2 by reactive oxygen and nitrogen species.

Cytokines such as TNF and FASL can trigger death or survival depending on cell lines and cellular conditions. The mechanistic details of how a cell chooses among these cell fates are still unclear. The understanding of these processes is important since they are altered in many diseases, including cancer and AIDS. Using a discrete modelling formalism, we present a mathematical model of cell fate decision recapitulating and integrating the most consistent facts extracted from the literature. This model provides a generic high-level view of the interplays between NFkappaB pro-survival pathway, RIP1-dependent necrosis, and the apoptosis pathway in response to death receptor-mediated signals. Wild type simulations demonstrate robust segregation of cellular responses to receptor engagement. Model simulations recapitulate documented phenotypes of protein knockdowns and enable the prediction of the effects of novel knockdowns. In silico experiments simulate the outcomes following ligand removal at different stages, and suggest experimental approaches to further validate and specialise the model for particular cell types. We also propose a reduced conceptual model implementing the logic of the decision process. This analysis gives specific predictions regarding cross-talks between the three pathways, as well as the transient role of RIP1 protein in necrosis, and confirms the phenotypes of novel perturbations. Our wild type and mutant simulations provide novel insights to restore apoptosis in defective cells. The model analysis expands our understanding of how cell fate decision is made. Moreover, our current model can be used to assess contradictory or controversial data from the literature. Ultimately, it constitutes a valuable reasoning tool to delineate novel experiments.

Cancerogenesis is driven by mutations leading to aberrant functioning of a complex network of molecular interactions and simultaneously affecting multiple cellular functions. Therefore, the successful application of bioinformatics and systems biology methods for analysis of high-throughput data in cancer research heavily depends on availability of global and detailed reconstructions of signalling networks amenable for computational analysis. We present here the Atlas of Cancer Signalling Network (ACSN), an interactive and comprehensive map of molecular mechanisms implicated in cancer. The resource includes tools for map navigation, visualization and analysis of molecular data in the context of signalling network maps. Constructing and updating ACSN involves careful manual curation of molecular biology literature and participation of experts in the corresponding fields. The cancer-oriented content of ACSN is completely original and covers major mechanisms involved in cancer progression, including DNA repair, cell survival, apoptosis, cell cycle, EMT and cell motility. Cell signalling mechanisms are depicted in detail, together creating a seamless 'geographic-like' map of molecular interactions frequently deregulated in cancer. The map is browsable using NaviCell web interface using the Google Maps engine and semantic zooming principle. The associated web-blog provides a forum for commenting and curating the ACSN content. ACSN allows uploading heterogeneous omics data from users on top of the maps for visualization and performing functional analyses. We suggest several scenarios for ACSN application in cancer research, particularly for visualizing high-throughput data, starting from small interfering RNA-based screening results or mutation frequencies to innovative ways of exploring transcriptomes and phosphoproteomes. Integration and analysis of these data in the context of ACSN may help interpret their biological significance and formulate mechanistic hypotheses. ACSN may also support patient stratification, prediction of treatment response and resistance to cancer drugs, as well as design of novel treatment strategies.

Thiol-based peroxidases consist of the peroxiredoxins (Prx) and the related glutathione peroxidase (GPx)-like enzymes. Their catalytic function is to reduce peroxides by using the reactivity of the cysteine residue, and their presumed primary physiologic role is to protect living organisms from peroxide toxicity. However, as peroxide-metabolizing enzymes, they also regulate hydrogen peroxide (H2O2) signaling. We review here enzymatic and biochemical attributes of thiol peroxidases that specify both distinctive peroxide-scavenging functions and the property of regulating H2O2 signaling. We then discuss possible thiol peroxidase physiologic functions, based on selected observations made in microorganisms and mammals.

Reactive oxygen species and nitric oxide (NO) are capable of both mediating redox-sensitive signal transduction and eliciting cell injury. The interplay between these messengers is quite complex, and intersection of their signaling pathways as well as regulation of their fluxes requires tight control. In this regard, peroxiredoxins (Prxs), a recently identified family of six thiol peroxidases, are central because they reduce H2O2, organic peroxides, and peroxynitrite. Here we provide evidence that endogenously produced NO participates in protection of murine primary macrophages against oxidative and nitrosative stress by inducing Prx I and VI expression at mRNA and protein levels. We also show that NO prevented the sulfinylation-dependent inactivation of 2-Cys Prxs, a reversible overoxidation that controls H2O2 signaling. In addition, studies using macrophages from sulfiredoxin (Srx)-deficient mice indicated that regeneration of 2-Cys Prxs to the active form was dependent on Srx. Last, we show that NO increased Srx expression and hastened Srx-dependent recovery of 2-Cys Prxs. We therefore propose that modulation by NO of Prx expression and redox state, as well as up-regulation of Srx expression, constitutes a novel pathway that contributes to antioxidant response and control of H2O2-mediated signal transduction in mammals. Reactive oxygen species and nitric oxide (NO) are capable of both mediating redox-sensitive signal transduction and eliciting cell injury. The interplay between these messengers is quite complex, and intersection of their signaling pathways as well as regulation of their fluxes requires tight control. In this regard, peroxiredoxins (Prxs), a recently identified family of six thiol peroxidases, are central because they reduce H2O2, organic peroxides, and peroxynitrite. Here we provide evidence that endogenously produced NO participates in protection of murine primary macrophages against oxidative and nitrosative stress by inducing Prx I and VI expression at mRNA and protein levels. We also show that NO prevented the sulfinylation-dependent inactivation of 2-Cys Prxs, a reversible overoxidation that controls H2O2 signaling. In addition, studies using macrophages from sulfiredoxin (Srx)-deficient mice indicated that regeneration of 2-Cys Prxs to the active form was dependent on Srx. Last, we show that NO increased Srx expression and hastened Srx-dependent recovery of 2-Cys Prxs. We therefore propose that modulation by NO of Prx expression and redox state, as well as up-regulation of Srx expression, constitutes a novel pathway that contributes to antioxidant response and control of H2O2-mediated signal transduction in mammals. Macrophages participate in many important functions, including phagocytosis, iron recycling, and host defense, and produce the autacoid-like reactive oxygen species (ROS) 3The abbreviations used are:ROSreactive oxygen speciesBMMbone marrow-derived macrophageNOnitric oxideNOS2nitric-oxide synthase 2DETA-NOdiethyltriamine-NONOatePMAphorbol 12-myristate 13-acetatePrxperoxiredoxinSrxsulfiredoxinLPSlipopolysaccharideIFNinterferonWTwild typeRT-PCRreverse transcription PCR. 3The abbreviations used are:ROSreactive oxygen speciesBMMbone marrow-derived macrophageNOnitric oxideNOS2nitric-oxide synthase 2DETA-NOdiethyltriamine-NONOatePMAphorbol 12-myristate 13-acetatePrxperoxiredoxinSrxsulfiredoxinLPSlipopolysaccharideIFNinterferonWTwild typeRT-PCRreverse transcription PCR. and nitric oxide (NO) in response to inflammatory cytokines and bacterial products. It has long been reported that ROS and reactive nitrogen species are effectors of an innate immune response (1Nathan C. Shiloh M.U. Proc. Natl. Acad. Sci. U. S. A. 2000; 97: 8841-8848Crossref PubMed Scopus (1148) Google Scholar), but there is increasing evidence that both ROS, and particularly H2O2, and NO also operate as signaling molecules to mediate various responses, including cell growth, angiogenesis, and apoptosis (2Forman H.J. Torres M. Mol. Aspects Med. 2001; 22: 189-216Crossref PubMed Scopus (437) Google Scholar, 3Nathan C. Sci. STKE. 2004; : pe52PubMed Google Scholar). Thus, H2O2 is now recognized as an important intracellular messenger that is physiologically produced by many cells in response to extracellular stimuli like cytokines and growth factors (4Rhee S.G. Kang S.W. Jeong W. Chang T.S. Yang K.S. Woo H.A. Curr. Opin. Cell Biol. 2005; 17: 183-189Crossref PubMed Scopus (608) Google Scholar). Second messenger functions mediated by H2O2 signaling include activation of mitogen-activated protein kinase (5Blanc A. Pandey N.R. Srivastava A.K. Int. J. Mol. Med. 2003; 11: 229-234Crossref PubMed Scopus (10) Google Scholar), modulation of the cell cycle (6Thomas D.D. Miranda K.M. Espey M.G. Citrin D. Jourd'heuil D. Paolocci N. Hewett S.J. Colton C.A. Grisham M.B. Feelisch M. Wink D.A. Methods Enzymol. 2002; 359: 84-105Crossref PubMed Scopus (73) Google Scholar, 7Phalen T.J. Weirather K. Deming P.B. Anathy V. Howe A.K. van der Vliet A. Jonsson T.J. Poole L.B. Heintz N.H. J. Cell Biol. 2006; 175: 779-789Crossref PubMed Scopus (117) Google Scholar), inhibition of tyrosine and lipid phosphatases (8Kwon J. Lee S.R. Yang K.S. Ahn Y. Kim Y.J. Stadtman E.R. Rhee S.G. Proc. Natl. Acad. Sci. U. S. A. 2004; 101: 16419-16424Crossref PubMed Scopus (511) Google Scholar, 9Leslie N.R. Bennett D. Lindsay Y.E. Stewart H. Gray A. Downes C.P. EMBO J. 2003; 22: 5501-5510Crossref PubMed Scopus (498) Google Scholar), and protein sumoylation (10Bossis G. Melchior F. Mol. Cell. 2006; 21: 349-357Abstract Full Text Full Text PDF PubMed Scopus (309) Google Scholar). Such signaling pathways imply a tight control of H2O2 production and elimination. reactive oxygen species bone marrow-derived macrophage nitric oxide nitric-oxide synthase 2 diethyltriamine-NONOate phorbol 12-myristate 13-acetate peroxiredoxin sulfiredoxin lipopolysaccharide interferon wild type reverse transcription PCR reactive oxygen species bone marrow-derived macrophage nitric oxide nitric-oxide synthase 2 diethyltriamine-NONOate phorbol 12-myristate 13-acetate peroxiredoxin sulfiredoxin lipopolysaccharide interferon wild type reverse transcription PCR Peroxiredoxins (Prxs) constitute an important peroxidase family that uses the reactivity of the cysteine residues to reduce H2O2 and other peroxides. Reaction of H2O2 with Prxs is fast as indicated by recent reassessment of the kinetic values (11Parsonage D. Youngblood D.S. Sarma G.N. Wood Z.A. Karplus P.A. Poole L.B. Biochemistry. 2005; 44: 10583-10592Crossref PubMed Scopus (172) Google Scholar, 12Peskin A.V. Low F.M. Paton L.N. Maghzal G.J. Hampton M.B. Winterbourn C.C. J. Biol. Chem. 2007; 282: 11885-11892Abstract Full Text Full Text PDF PubMed Scopus (311) Google Scholar). Further, in addition to their antioxidant function, Prxs have been shown to regulate cell signaling by H2O2 by modulating its fluxes and intracellular levels (13Lee S.R. Kwon K.S. Kim S.R. Rhee S.G. J. Biol. Chem. 1998; 273: 15366-15372Abstract Full Text Full Text PDF PubMed Scopus (840) Google Scholar, 14Cho S.H. Lee C.H. Ahn Y. Kim H. Kim H. Ahn C.Y. Yang K.S. Lee S.R. FEBS Lett. 2004; 560: 7-13Crossref PubMed Scopus (171) Google Scholar). It is also worth noting that Prxs can reduce peroxynitrite (15Bryk R. Griffin P. Nathan C. Nature. 2000; 407: 211-215Crossref PubMed Scopus (563) Google Scholar, 16Rhee S.G. Chae H.Z. Kim K. Free Radic. Biol. Med. 2005; 38: 1543-1552Crossref PubMed Scopus (1135) Google Scholar). Mammals carry six Prx enzymes that distribute in the three Prx subtypes with four typical 2-Cys Prxs (I-IV), one atypical 2-Cys Prx (Prx V), and one 1-Cys Prx (Prx VI) (17Wood Z.A. Schroder E. Robin Harris J. Poole L.B. Trends Biochem. Sci. 2003; 28: 32-40Abstract Full Text Full Text PDF PubMed Scopus (2100) Google Scholar). Typical 2-Cys Prxs have the unique feature of undergoing substrate-mediated inactivation by overoxidation of their catalytic cysteine to a sulfinic acid (R-SO2H). Overoxidation only occurs during enzymatic cycling and is proportional to the amount of substrate under both non-saturating and saturating conditions (15Bryk R. Griffin P. Nathan C. Nature. 2000; 407: 211-215Crossref PubMed Scopus (563) Google Scholar). The fact that inactivation by overoxidation is both unique to eukaryotic Prxs and reversible by ATP-dependent reduction of the Prx Cys-SO2H by sulfiredoxin (Srx or npn3) and sestrins (18Biteau B. Labarre J. Toledano M.B. Nature. 2003; 425: 980-984Crossref PubMed Scopus (793) Google Scholar, 19Woo H.A. Kang S.W. Kim H.K. Yang K.S. Chae H.Z. Rhee S.G. J. Biol. Chem. 2003; 278: 47361-47364Abstract Full Text Full Text PDF PubMed Scopus (200) Google Scholar, 20Chevallet M. Wagner E. Luche S. van Dorsselaer A. Leize-Wagner E. Rabilloud T. J. Biol. Chem. 2003; 278: 37146-37153Abstract Full Text Full Text PDF PubMed Scopus (146) Google Scholar, 21Chang T.S. Jeong W. Woo H.A. Lee S.M. Park S. Rhee S.G. J. Biol. Chem. 2004; 279: 50994-51001Abstract Full Text Full Text PDF PubMed Scopus (306) Google Scholar, 22Budanov A.V. Sablina A.A. Feinstein E. Koonin E.V. Chumakov P.M. Science. 2004; 304: 596-600Crossref PubMed Scopus (615) Google Scholar) had led to the suggestion that it is an acquired gain of function selected for regulating intracellular H2O2 fluxes and signaling (23Wood Z.A. Poole L.B. Karplus P.A. Science. 2003; 300: 650-653Crossref PubMed Scopus (1135) Google Scholar). Hence, 2-Cys Prx activity is controlled both by the levels of its substrate H2O2 and by the activity of sulfinyl reductases, and this dual control is likely important for regulating H2O2 signaling. In this report, we have investigated the impact of NO on the expression of Prxs, Srx, and sestrins in murine macrophages. We provide a global view of the expression of the six mammalian Prxs in macrophages that produce NO upon stimulation with interferon γ (IFN-γ) and lipopolysaccharide (LPS). We show that gene expression of Prx I, V, and VI and of Srx was increased in stimulated macrophages. Up-regulation of Prx I and VI, but not Prx V, was mediated by NO. We also report that NO decreases spontaneous and H2O2-induced Prx sulfinylation and hastens recovery upon H2O2-induced Prx sulfinylation, thus pointing to a role for NO in overoxidation prevention and reactivation of 2-Cys Prx. Reagents—Recombinant mouse IFN-γ (specific activity 2 × 106 units/mg) was from R&D Systems, Abigdon, UK). Escherichia coli LPS, N-(3-(aminomethyl-benzyl-acetamidine)) (1400W), phorbol 12-myristate 13-acetate (PMA), tert-butyl hydroperoxide, and cycloheximide were from Sigma. S-ethylisothiourea and the nitric oxide donor diethylenetriamine NONOate (DETA-NO) were from Cayman Chemical (Ann Arbor, MI). Glucose oxidase was from Calbiochem. Cell Culture and Treatments—Protocols involving animal experimentation were approved by a national animal care committee. Bone marrow cells were obtained by flushing femurs of WT C57BL/6 mice and of NOS2–/– or Srx–/– mice. Bone marrow-derived macrophages (BMM) were differentiated from bone marrow cells by culture in RPMI 1640 (Invitrogen) supplemented with 10% fetal bovine serum (Invitrogen) and 10% L929 cell-conditioned medium. The of was by cell It was shown that of the cells the were stimulated or not with IFN-γ E. coli at the and for the indicated in the were with the nitric oxide donor was by the of the at of Cell cells were with and in in Cell was at × at for and the protein of was at by using the protein and I was from was from and to Prx Prx VI, Prx and Prx were from The was a serum by was from Cell were by in or under the and protein were with and with primary were with peroxidase using or to or and was from cells using the to the of was using the murine reverse and PCR was using a and the of was using the I The of PCR was by were with was to the were in the of was in culture by using the of were with of and in and the was at The was from a H2O2 of H2O2 was using the were in a and for with in were stimulated for and with was with a by at NO Prx I and Prx VI in from WT or NOS2–/– mice were stimulated with IFN-γ and that had in the culture was and the expression of the six Prxs was by In stimulated WT Prx I, Prx V, and Prx VI mRNA expression was increased with Prx was In in stimulated NOS2–/– mRNA of Prx I, Prx and Prx VI with stimulated WT that regulation was dependent on Prx mRNA levels increased in NOS2–/– that the up-regulation of this gene by IFN-γ and is of Prx and Prx mRNA levels were not by stimulation in mouse We also used an NO donor with a long that NO at in the produced by IFN-γ and macrophages S. R. G. Proc. Natl. Acad. Sci. U. S. A. 2001; PubMed Scopus Google Scholar). of macrophages during the of NO produced by on Prx gene to that Prx VI and Prx I mRNA levels increased and and that regulation is at in the of that had been to for at had on gene expression, that is for the regulation of Prxs with these to a role of NO in the regulation of Prx I, and VI mRNA levels. We the in Prx I and VI levels also at protein Prx I and VI protein levels were in stimulated with IFN-γ and LPS, but not cells were stimulated in the of or using cells from mice Prx I and Prx VI protein levels were also increased in with in a In Prx and protein levels were not to NO the these the role of NO in up-regulation of Prx I and VI protein of NO on Prx active cysteine of eukaryotic Prxs substrate-mediated to the sulfinic acid form activity T. M. F. Luche S. C. R. M. P. J. J. Biol. Chem. 2002; Full Text Full Text PDF PubMed Scopus Google Scholar, K.S. Kang S.W. Woo H.A. Chae H.Z. Kim K. Rhee S.G. J. Biol. Chem. 2002; Full Text Full Text PDF PubMed Scopus Google Scholar). on the NO cysteine sulfinylation in K. M. Karplus P.A. Biol. 1998; PubMed Scopus Google Scholar, T. M. K. M. N. Biochemistry. 2003; PubMed Scopus Google Scholar), we NO also overoxidation of Prxs. an that with the form of the four 2-Cys Prxs we thus the of Prxs in macrophages that were stimulated to produce NO In of macrophages used as a the the of and one on and H.A. Kang S.W. Kim H.K. Yang K.S. Chae H.Z. Rhee S.G. J. Biol. Chem. 2003; 278: 47361-47364Abstract Full Text Full Text PDF PubMed Scopus (200) Google Scholar, H.A. Jeong W. Chang T.S. Park Park S.J. Yang Rhee S.G. J. Biol. Chem. 2005; Full Text Full Text PDF PubMed Scopus Google Scholar), the is Prx and the to Prx the is Prx In control WT a unique was at the of Prx is to the of ROS production by macrophages. in and Prx overoxidation was not or but in macrophages and in macrophages stimulated in the of overoxidation was at the levels of the species for the in Prx overoxidation in stimulated we the of of to also Prx overoxidation was at as as and was increased at indicated that the of NO on Prx with an at 2 addition of We NO also the overoxidation of Prxs by were with IFN-γ and or were to for they were with a of H2O2 with the or with H2O2 produced at by the oxidase H2O2-induced Prx Prx to a Prx overoxidation was in macrophages as with the with to Prx were obtained with cells to NO pointing to a on various cell and We also used a that H2O2 with led to H2O2 as by of the also increased the levels of 2-Cys Prxs, was not cells had been with We also 1-Cys Prx VI overoxidation was also by using a Prx Prx VI was only of to H2O2, and stimulation of by or to not Prx VI overoxidation Prx VI protein levels were increased by NO as shown and with an VI Prx was not because this Prx is not to overoxidation M. Wagner E. Luche S. van Dorsselaer A. Leize-Wagner E. Rabilloud T. J. Biol. Chem. 2003; 278: 37146-37153Abstract Full Text Full Text PDF PubMed Scopus (146) Google Scholar). In these that NO decreases 2-Cys Prx sulfinylation this form of by NO Prx sulfinylation or its on the of NO on up-regulation of Prx I expression, it is that the increased as a Prx I the fact that the cycloheximide not from H2O2-mediated Prx overoxidation that Prx I up-regulation and overoxidation can also show that in Prx overoxidation not protein of NO on Srx 2-Cys Prx can by of enzymes with ATP-dependent sulfinic acid Srx (18Biteau B. Labarre J. Toledano M.B. Nature. 2003; 425: 980-984Crossref PubMed Scopus (793) Google Scholar, 21Chang T.S. Jeong W. Woo H.A. Lee S.M. Park S. Rhee S.G. J. Biol. Chem. 2004; 279: 50994-51001Abstract Full Text Full Text PDF PubMed Scopus (306) Google Scholar) and the sestrins A.V. Sablina A.A. Feinstein E. Koonin E.V. Chumakov P.M. Science. 2004; 304: 596-600Crossref PubMed Scopus (615) Google Scholar). We therefore investigated the of Srx and sestrins in the in Prx mRNA levels were not in or In Srx mRNA levels were in macrophages and this was dependent on NO because it was not in stimulated macrophages from NOS2–/– mice. that the in Srx expression in as as and using an that the in Srx mRNA levels was by an in Srx protein levels We to the up-regulation of Srx and the in Prx overoxidation were this we the Prx in from WT or mice that had been to and with shown in of WT with both and H2O2-induced Prx overoxidation with and with The were in Srx–/– with and with that the of NO on the of Prx overoxidation not Srx. a role of Srx on Prx we the of Prx sulfinic acid a to Prx sulfinylation was at in with that were to H2O2 during and and in of that the of the H2O2-induced sulfinic acid signal had by in WT control and only in WT in of Srx–/– that had been or not with the signal of H2O2-induced Prx not the that NO not only Prx sulfinylation but also the of its in also the role of Srx in the reduction of 2-Cys Prxs in mammals. by and species is as its in is T. Curr. Opin. Cell Biol. 1998; PubMed Scopus Google Scholar, H.J. Torres M. J. Mol. Cell. Biochem. 2002; PubMed Scopus Google Scholar). these are they can also various their control of for cell (1Nathan C. Shiloh M.U. Proc. Natl. Acad. Sci. U. S. A. 2000; 97: 8841-8848Crossref PubMed Scopus (1148) Google Scholar, M.G. Miranda K.M. D.D. S. Citrin D. Wink D.A. N. Y. Acad. Sci. 2002; PubMed Scopus Google Scholar, D.D. Espey M.G. S. S. Harris C.C. W. D.D. Wink D.A. J. Biol. Chem. 2006; Full Text Full Text PDF PubMed Scopus Google Scholar). Prxs are important enzymes and have been shown to also signaling by 2-Cys Prxs, Prx reversible inactivation by overoxidation of their catalytic cysteine to the sulfinic acid form S.G. Chae H.Z. Kim K. Free Radic. Biol. Med. 2005; 38: 1543-1552Crossref PubMed Scopus (1135) Google Scholar, Z.A. Poole L.B. Karplus P.A. Science. 2003; 300: 650-653Crossref PubMed Scopus (1135) Google Scholar) and reduction by ATP-dependent sulfiredoxin and sestrins (18Biteau B. Labarre J. Toledano M.B. Nature. 2003; 425: 980-984Crossref PubMed Scopus (793) Google Scholar, 21Chang T.S. Jeong W. Woo H.A. Lee S.M. Park S. Rhee S.G. J. Biol. Chem. 2004; 279: 50994-51001Abstract Full Text Full Text PDF PubMed Scopus (306) Google Scholar). unique to eukaryotic Prxs, has led to that it is an acquired gain of function selected for (23Wood Z.A. Poole L.B. Karplus P.A. Science. 2003; 300: 650-653Crossref PubMed Scopus (1135) Google Scholar). In the we have identified a novel between the of NO and H2O2 in primary macrophages. is on the of NO of H2O2-induced 2-Cys Prx sulfinylation, of the regeneration of and of Prx I, Prx VI, and Srx at mRNA and protein levels. were NO was endogenously produced by or by the NO donor and Prxs were by H2O2 that was endogenously produced by stimulated macrophages or also provide evidence that Prx is in macrophages stimulated by IFN-γ and LPS, but in to Prx I and Prx VI this is of NO in to was for 2-Cys Prxs, H2O2-mediated overoxidation of 1-Cys Prx VI was not by or is with the fact that Srx is not to reduce VI H.A. Jeong W. Chang T.S. Park Park S.J. Yang Rhee S.G. J. Biol. Chem. 2005; Full Text Full Text PDF PubMed Scopus Google Scholar). these that NO production the antioxidant of both by increasing Prx I and VI levels and by typical 2-Cys Prxs in their active particularly macrophages of are in with the of evidence that NO has antioxidant D.D. Espey M.G. S. S. Harris C.C. W. D.D. Wink D.A. J. Biol. Chem. 2006; Full Text Full Text PDF PubMed Scopus Google Scholar, A. P. E.R. J. Biol. Chem. Full Text Full Text PDF PubMed Scopus Google Scholar, D.A. W. J. Proc. Natl. Acad. Sci. U. S. A. PubMed Scopus Google Scholar, J. C. E. 2006; PubMed Scopus Google Scholar) protection against the of H2O2 J. S. R. Biochem. PubMed Scopus Google Scholar, P. P. 38: PubMed Scopus Google Scholar). they provide a of this as in the of Prx is also as an peroxynitrite M. D. A. F. T. R. B. FEBS Lett. 2004; PubMed Scopus Google Scholar) and is in a to a of peroxynitrite production S. J. 2006; 97: PubMed Scopus Google Scholar). the increased expression of Prx in and macrophages also a protection for the of NO that is produced by these We thus propose that stimulated macrophages are likely to from a of active Prx to the of and nitrosative but up-regulation of Prx I had been in macrophages S. J. S. G. PubMed Scopus Google Scholar), but the of this regulation was not The Prx I and Prx VI for the transcription Free Radic. Biol. Med. 2007; PubMed Scopus Google Scholar, Y.J. Ahn P. C. Y. Park 2007; PubMed Scopus Google Scholar, Kim S.G. Methods Enzymol. 2005; PubMed Scopus Google Scholar), and macrophages of mice are to Prx I expression in response to H2O2 and to T. K. S. H. T. Y. S. M. J. Biol. Chem. 2000; Full Text Full Text PDF PubMed Scopus Google Scholar). Srx to as was in a in the of mice with the as shown by N. K. H. M. J. Biol. Chem. 2003; 278: Full Text Full Text PDF PubMed Scopus Google Scholar). it is well that NO from NO including antioxidant response gene expression in mammalian cells Free Radic. Biol. Med. 2007; PubMed Scopus Google Scholar, T. K. S. H. T. Y. S. M. J. Biol. Chem. 2000; Full Text Full Text PDF PubMed Scopus Google Scholar, S. J. Biol. Chem. 2004; 279: Full Text Full Text PDF PubMed Scopus (146) Google Scholar). It is therefore to propose that the contributes to an response to oxidative and nitrosative by up-regulation of these three redox The NO 2-Cys Prx that NO both prevented catalytic cysteine overoxidation and the of its recovery by sulfiredoxin (18Biteau B. Labarre J. Toledano M.B. Nature. 2003; 425: 980-984Crossref PubMed Scopus (793) Google Scholar, 21Chang T.S. Jeong W. Woo H.A. Lee S.M. Park S. Rhee S.G. J. Biol. Chem. 2004; 279: 50994-51001Abstract Full Text Full Text PDF PubMed Scopus (306) Google Scholar). We that WT of this was as as 2 cells to NO it not the of reduction of the form of Prxs by Srx because of the gene by NO at We also that the overoxidation of Prxs that from a to H2O2 was in also that the not a of a reduction by Srx. In addition, the of NO on the overoxidation of Prxs by H2O2 was in Srx–/– that the of NO in 2-Cys Prx overoxidation is to Srx of are produced by stimulated have recently been shown to 2-Cys Prxs in or cell P. K. K. J. Biol. Chem. 2007; 282: Full Text Full Text PDF PubMed Scopus Google Scholar). It worth that by lipid N. B. PubMed Scopus Google Scholar, A. Biol. 2000; PubMed Scopus Google Scholar), decreases the of 2-Cys Prx a is that Prxs from macrophages are from overoxidation by H2O2 by a of its catalytic cysteine on the role of in the response to oxidative stress P. S. J. Biochem. 2000; PubMed Scopus Google Scholar, M.B. A. J. 2006; PubMed Scopus Google Scholar), protection of the Prx active by with is an NO also the of the Srx-dependent of Prxs. was from the in the amount of by H2O2 in macrophages also that in H2O2-induced Prx sulfinylation that Srx is the macrophage sulfinyl and that other can for its in these is in with the of Chang T.S. Jeong W. Woo H.A. Lee S.M. Park S. Rhee S.G. J. Biol. Chem. 2004; 279: 50994-51001Abstract Full Text Full Text PDF PubMed Scopus (306) Google Scholar) that Srx Prx regeneration in It also the that the of NO in Prx regeneration is mediated by Srx thus the role of Srx in modulating the redox of Prxs in a by pointing to Prxs as a between NO and H2O2 we propose a novel control by physiologically produced NO an antioxidant in the amount of active Prxs is likely to a to against It also to the redox control of the Further, the of the and as a in the of H2O2 a for of a between host and cell redox signaling. We C. H. and for with NOS2–/– were by C. Nathan of and J. and M. and P. for the of the Srx–/– mice. We are also to J. for cell and J. for with