title: Multimodal control of Cdc25A by nitrosative stress.
abstract: Cdc25A propels cell cycle progression, is overexpressed in numerous human cancers, and possesses oncogenic and antiapoptotic activities. Reactive oxygen species, such as hydrogen peroxide, regulate Cdc25A, but the physiologic and pathologic effects of nitric oxide (*NO) and *NO-derived reactive species are not well defined. Herein, we report novel independent mechanisms governing Cdc25A in response to nitrosative insult. We observed direct and rapid inhibition of Cdc25A phosphatase activity after in vitro treatment with the low molecular mass cell-permeable S-nitrosothiol S-nitrosocysteine ethyl ester (SNCEE). In addition, treatment of cancer cells with SNCEE induced nitrosative stress and decreased Cdc25A protein levels in a time-dependent and concentration-dependent manner. Similarly, iNOS-derived *NO was sufficient to suppress Cdc25A expression, consistent with its role in mediating nitrosative stress. Whereas a decrease in Cdc25A half-life was not observed in response to SNCEE, we found the translational regulator eukaryotic initiation factor 2alpha (eIF2alpha) was hyperphosphorylated and total protein translation was decreased with kinetics consistent with Cdc25A loss. Inhibition of eIF2alpha decreased Cdc25A levels, supporting the hypothesis that SNCEE suppressed Cdc25A translation through inhibition of eIF2alpha. Nitrosative stress decreased the Cdc25A-bound fraction of apoptosis signal-regulating kinase-1 (ASK-1) and sensitized cells to apoptosis induced by the ASK-1-activating chemotherapeutic cis-diaminedichloroplatinum (II), suggesting that nitrosative stress-induced suppression of Cdc25A primed cells for ASK-1-dependent apoptosis. Together these data reveal novel *NO-dependent enzymatic and translational mechanisms controlling Cdc25A, and implicate Cdc25A as a mediator of *NO-dependent apoptotic signaling.
authors.affiliation: Department of Pharmacology and Chemical Biology and University of Pittsburgh Drug Discovery Institute, University of Pittsburgh, Pittsburgh, Pennsylvania 15260, USA.
authors.forename: Robert J
authors.initials: RJ
authors.lastname: Tomko
authors.forename: John S
authors.initials: JS
authors.lastname: Lazo
mesh_terms: D017209:Apoptosis / Q000187:drug effects / Q000502:physiology; D045744:Cell Line, Tumor; D002945:Cisplatin / Q000494:pharmacology; D003545:Cysteine / Q000031:analogs & derivatives / Q000494:pharmacology; D015536:Down-Regulation; D015852:Eukaryotic Initiation Factor-2 / Q000037:antagonists & inhibitors / Q000378:metabolism; D045325:HCT116 Cells; D006801:Humans; D007700:Kinetics; D048848:MAP Kinase Kinase Kinase 5 / Q000378:metabolism; D009569:Nitric Oxide / Q000378:metabolism*; D052247:Nitric Oxide Synthase Type II / Q000378:metabolism; D009603:Nitroso Compounds / Q000494:pharmacology; D010084:Oxidation-Reduction; D010766:Phosphorylation; D014176:Protein Biosynthesis; D020687:cdc25 Phosphatases / Q000037:antagonists & inhibitors / Q000096:biosynthesis / Q000235:genetics / Q000378:metabolism*
chemical_list: D015852:Eukaryotic Initiation Factor-2; D009603:Nitroso Compounds; C429310:S-nitrosocysteine ethyl ester; D009569:Nitric Oxide; C496319:NOS2 protein, human; D052247:Nitric Oxide Synthase Type II; D048848:MAP Kinase Kinase Kinase 5; C482858:MAP3K5 protein, human; C517004:CDC25A protein, human; D020687:cdc25 Phosphatases; D003545:Cysteine; D002945:Cisplatin
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