S-Nitrosoglutathione and Endothelial Nitric Oxide Synthase-Derived Nitric Oxide Regulate Compartmentalized Ras S-Nitrosylation and Stimulate Cell Proliferation

Autor(es)

Batista, Wagner L. ^[1] ; Ogata, Fernando T. ^[2] ; Curcio, Marli F. ^[3] ; Miguel, Rodrigo B. ^[2] ; Arai, Roberto J. ^[4] ; Matsuo, Alisson L. ^[3] ; Moraes, Miriam S. ^[2] ; Stern, Arnold ^[5] ; Monteiro, Hugo P. ^[2]

Afiliação do(s) autor(es)

^[1] Univ Fed Sao Paulo, Dept Biol Sci, BR-04044010 Sao Paulo - Brazil

^[2] Univ Fed Sao Paulo, CTCMOL, Dept Biochem Mol Biol, BR-04044010 Sao Paulo - Brazil

^[3] Univ Fed Sao Paulo, Dept Microbiol Parasitol & Immunol, BR-04044010 Sao Paulo - Brazil

^[4] Univ Sao Paulo, Sch Med, Sao Paulo Canc Inst, Sao Paulo - Brazil

^[5] NYU, Sch Med, Dept Pharmacol, New York, NY - USA

• Tipo de documento: Artigo Científico
• Fonte: Antioxidants & Redox Signaling; v. 18, n. 3, p. 221-238, JAN 2013.
• Citações Web of Science: 23

Resumo

Aims: S-nitrosylation of Cys118 is a redox-based mechanism for Ras activation mediated by nitric oxide (NO) at the plasma membrane. Results: Ras signaling pathway stimulation by 50 and/or 100 mu M of S-nitrosoglutathione (GSNO) causes proliferation of HeLa cells. Proliferation was not observed in HeLa cells overexpressing non-nitrosatable H-Ras(C118S). HeLa cells overexpressing H-Ras(wt) containing the spatiotemporal probe green fluorescent protein (GFP) fused to the Ras-binding domain of Raf-1 (GFP-RBD) incubated with 100 mu M GSNO stimulated a rapid and transient redistribution of GFP-RBD to the plasma membrane, followed by a delayed and sustained recruitment to the Golgi. No activation of H-Ras at the plasma membrane occurred in cells overexpressing H-Ras(C118S), contrasting with a robust and sustained activation of the GTPase at the Golgi. Inhibition of Src kinase prevented cell proliferation and activation of H-Ras by GSNO at the Golgi. Human umbilical vein endothelial cells (HUVECs) stimulated with bradykinin to generate NO were used to differentiate cell proliferation and Ras activation at the plasma membrane versus Golgi. In this model, Src kinase was not involved in cell proliferation, whereas Ras activation proceeded only at the plasma membrane, indicating that HUVEC proliferation induced by NO resulted only from stimulation of Ras. Innovation: The present work is the first to demonstrate that NO-mediated activation of Ras in different subcellular compartments regulates different downstream signaling pathways. Conclusion: S-nitrosylation of H-Ras at Cys(118) and the activation of Src kinase are spatiotemporally linked events of the S-nitrosothiol-mediated signaling pathway that occurs at the plasma membrane and at the Golgi. The nonparticipation of Src kinase and the localized production of NO by endothelial NO synthase at the plasma membrane limited NO-mediated Ras activation to the plasma membrane. Antioxid. Redox Signal. 18, 221-238. (AU)

• Processo FAPESP: 11/14392-2 - Avaliação do envolvimento da GTPase Ras de Paracoccidioides brasiliensis no termo-dimorfismo do fungo e durante o estresse oxidativo e nitrosativo
• Beneficiário: Wagner Luiz Batista
• Modalidade de apoio: Auxílio à Pesquisa - Regular
• Processo FAPESP: 09/52730-7 - Estudo do papel do oxido nitrico na ativacao da gtpase rac-1 e seu envolvimento na manutencao e progressao da metastase tumoral em modelo de melanoma murino.
• Beneficiário: Hugo Pequeno Monteiro
• Modalidade de apoio: Auxílio à Pesquisa - Regular
• Processo FAPESP: 07/59617-6 - Estudos da compartimentalização celular da proteína G de baixo peso molecular Ras e da proteína antioxidante tiorredoxina sob estímulo de fontes geradoras de óxido nítrico
• Beneficiário: Hugo Pequeno Monteiro
• Modalidade de apoio: Auxílio à Pesquisa - Regular