Analysis of the nitric oxide production pathway in venous endothelial cells from rats
Role of syndecan-4 in extracellular matrix remodeling in anoikis-resistant endothe...
| Full text | |
| Author(s): |
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]
Total Authors: 9
|
| Affiliation: | [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
Total Affiliations: 5
|
| Document type: | Journal article |
| Source: | Antioxidants & Redox Signaling; v. 18, n. 3, p. 221-238, JAN 2013. |
| Web of Science Citations: | 23 |
| Abstract | |
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) | |
| FAPESP's process: | 11/14392-2 - Evaluation of the involvement of Ras GTPase from Paracoccidioides brasiliensis in thermo-dimorphism and during oxidative and nitrosative stress |
| Grantee: | Wagner Luiz Batista |
| Support Opportunities: | Regular Research Grants |