title: S-nitrosylation of Cofilin-1 Serves as a Novel Pathway for VEGF-Stimulated Endothelial Cell Migration.
abstract: Nitric oxide (NO) derived from endothelial NO synthase (eNOS) mediates vascular endothelial growth factor (VEGF)-stimulated endothelial cytoskeleton remodeling and migration; however, the underlying mechanisms are elusive. Covalent adduction of a NO moiety (NO(•)) to cysteines called S-nitrosylation (SNO) is a key NO signaling pathway. The small actin-binding protein cofilin-1 (CFL1) is essential for actin cytoskeleton remodeling. We investigated whether S-nitrosylation regulates CFL1 function and endothelial cytoskeleton remodeling and migration upon VEGF stimulation. VEGF rapidly stimulated S-nitrosylation of CFL1, which was blocked by NO Synthase inhibition and eNOS knockdown by specific eNOS-siRNA. Cys80 and Cys139 were identified as the major SNO-sites in CFL1 by LC-MS/MS. The actin severing activity of recombinant SNO-mimetic CFL1 (C80/139A DMA-CFL1), but not SNO-deficient CFL1 (C80/139S DMS-CFL1), was significantly greater than that of wild-type CFL1 (wt-CFL1). When wt-CFL1 and its mutants were overexpressed in endothelial cells, basal actin bound wt-CFL1 was undetectable but significantly increased by VEGF; basal actin bound DMA-CFL1 was readily high and basal actin bound DMS-CFL1 was detectable but low, and both were unresponsive to VEGF. Treatment with VEGF significantly increased filamentous (F-) actin and filopodium formation and cell migration in endothelial cells. Overexpression of wt-CFL1 inhibited VEGF-induced F-actin formation. Overexpression of DMA but not DMS CFL1 decreased basal but not VEGF-stimulated F-actin formation. Overexpression of DMA but not DMS CFL1 suppressed VEGF-stimulated filopodium formation and migration in endothelial cells. Thus, S-nitrosylation of CFL1 provides a novel signaling pathway post-NO biosynthesis via eNOS-derived NO for endothelial cytoskeleton remodeling and migration upon VEGF stimulation.
authors: Department of Obstetrics and Gynecology, University of California, Irvine, California.
authors: Hong-Hai
authors: HH
authors: Zhang
authors: Department of Obstetrics and Gynecology, University of California, Irvine, California.
authors: Wen
authors: W
authors: Wang
authors: Department of Obstetrics and Gynecology, University of California, Irvine, California.
authors: Lin
authors: L
authors: Feng
authors: Department of Biophysics and Physiology, University of California, Irvine, California.
authors: Yingying
authors: Y
authors: Yang
authors: Department of Obstetrics and Gynecology, University of Wisconsin-Madison, Madison, Wisconsin.
authors: Jing
authors: J
authors: Zheng
authors: Department of Biophysics and Physiology, University of California, Irvine, California.
authors: Lan
authors: L
authors: Huang
authors: Department of Obstetrics and Gynecology, University of California, Irvine, California.
authors: Dong-Bao
authors: DB
authors: Chen
mesh_terms: D000199:Actins / Q000378:metabolism; D002465:Cell Movement; D002478:Cells, Cultured; D051338:Cofilin 1 / Q000378:metabolism*; D003599:Cytoskeleton / Q000378:metabolism*; D042783:Endothelial Cells / Q000378:metabolism*; D006801:Humans; D008840:Microfilament Proteins / Q000378:metabolism; D052250:Nitric Oxide Synthase Type III / Q000378:metabolism; D015398:Signal Transduction; D053719:Tandem Mass Spectrometry / Q000379:methods; D042461:Vascular Endothelial Growth Factor A / Q000378:metabolism*
chemical_list: D000199:Actins; D051338:Cofilin 1; D008840:Microfilament Proteins; D042461:Vascular Endothelial Growth Factor A; D052250:Nitric Oxide Synthase Type III