Vimentin expression influences flow dependent VASP phosphorylation and regulates cell migration and proliferation

TY - JOUR

T1 - Vimentin expression influences flow dependent VASP phosphorylation and regulates cell migration and proliferation

AU - Lund, Natalie

AU - Henrion, Daniel

AU - Tiede, Petra

AU - Ziche, Marina

AU - Schunkert, Heribert

AU - Ito, Wulf D

N1 - Copyright (c) 2010 Elsevier Inc. All rights reserved.

PY - 2010/5/7

Y1 - 2010/5/7

N2 - The cytoskeleton plays a central role for the integration of biochemical and biomechanical signals across the cell required for complex cellular functions. Recent studies indicate that the intermediate filament vimentin is necessary for endothelial cell morphogenesis e.g. in the context of leukocyte transmigration. Here, we present evidence, that the scaffold provided by vimentin is essential for VASP localization and PKG mediated VASP phosphorylation and thus controls endothelial cell migration and proliferation. Vimentin suppression using siRNA technique significantly decreased migration velocity by 50% (videomicroscopy), diminished transmigration activity by 42.5% (Boyden chamber) and reduced proliferation by 43% (BrdU-incorporation). In confocal microscopy Vimentin colocalized with VASP and PKG in endothelial cells. Vimentin suppression was accompanied with a translocation of VASP from focal contacts to the perinuclear region. VASP/Vimentin and PKG/Vimentin colocalization appeared to be essential for proper PKG mediated VASP phosphorylation because we detected a diminished expression of PKG and p(Ser239)-VASP in vimentin-suppressed cells, Furthermore, the induction of VASP phosphorylation in perfused arteries was markedly decreased in vimentin knockout mice compared to wildtypes. A link is proposed between vimentin, VASP phosphorylation and actin dynamics that delivers an explanation for the important role of vimentin in controlling endothelial cell morphogenesis.

AB - The cytoskeleton plays a central role for the integration of biochemical and biomechanical signals across the cell required for complex cellular functions. Recent studies indicate that the intermediate filament vimentin is necessary for endothelial cell morphogenesis e.g. in the context of leukocyte transmigration. Here, we present evidence, that the scaffold provided by vimentin is essential for VASP localization and PKG mediated VASP phosphorylation and thus controls endothelial cell migration and proliferation. Vimentin suppression using siRNA technique significantly decreased migration velocity by 50% (videomicroscopy), diminished transmigration activity by 42.5% (Boyden chamber) and reduced proliferation by 43% (BrdU-incorporation). In confocal microscopy Vimentin colocalized with VASP and PKG in endothelial cells. Vimentin suppression was accompanied with a translocation of VASP from focal contacts to the perinuclear region. VASP/Vimentin and PKG/Vimentin colocalization appeared to be essential for proper PKG mediated VASP phosphorylation because we detected a diminished expression of PKG and p(Ser239)-VASP in vimentin-suppressed cells, Furthermore, the induction of VASP phosphorylation in perfused arteries was markedly decreased in vimentin knockout mice compared to wildtypes. A link is proposed between vimentin, VASP phosphorylation and actin dynamics that delivers an explanation for the important role of vimentin in controlling endothelial cell morphogenesis.

KW - Animals

KW - Cell Adhesion Molecules/metabolism

KW - Cell Movement

KW - Cell Proliferation

KW - Cyclic GMP-Dependent Protein Kinases/metabolism

KW - Endothelial Cells/cytology

KW - Mice

KW - Mice, Knockout

KW - Microfilament Proteins/metabolism

KW - Phosphoproteins/metabolism

KW - Phosphorylation

KW - Rats

KW - Serine/metabolism

KW - Vimentin/biosynthesis

U2 - 10.1016/j.bbrc.2010.04.033

DO - 10.1016/j.bbrc.2010.04.033

M3 - SCORING: Journal article

C2 - 20382123

VL - 395

SP - 401

EP - 406

JO - BIOCHEM BIOPH RES CO

JF - BIOCHEM BIOPH RES CO

SN - 0006-291X

IS - 3

ER -