title: Subcellular localization of RhoA and ezrin at membrane ruffles of human endothelial cells: differential role of collagen and fibronectin.
abstract: Endothelial cells and the regulation of their migration are of prime importance in many physiological and pathological processes such as angiogenesis. RhoA, an important Rho family member known to trigger actin reorganization, has been shown to mediate the formation of focal adhesions and stress fibers in quiescent fibroblasts. However, recent studies have emphasized its functional diversity and its implication in migration or metastatic processes in different cell types other than fibroblasts. Its role in endothelial cells is little known. In this study, we were interested by analyzing in human endothelial cells the subcellular redistribution of endogenous RhoA and the reorganization of cytoskeletal actin induced by two important extracellular matrix proteins, collagen and fibronectin. This paper shows a translocation of RhoA and its association with cortical actin in focal contact domains at membrane ruffles and at lamellipodia of spread or migrating endothelial cells, in the absence of any soluble mitogen stimulation. Furthermore, RhoA was found colocalized with ezrin, a member of the ERM family proteins newly described as important membrane-actin cytoskeleton linkers, at early membrane ruffles of endothelial cells spread on collagen but not on fibronectin. The present study points out that extracellular matrix, depending on the nature of its components, may promote distinct assemblies of focal contact constitutive proteins and strongly suggests that endothelial RhoA, like Rac1, may be an important mediator of matrix signaling pathway regulating endothelial cell adhesiveness and motility, independently of growth factor stimulation.
authors.affiliation: Institut National de la Santé et de la Recherche Medicale U353, Laboratoire d'Analyse d'Images en Pathologie Cellulaire, Service d'Imagerie Cellulaire et Moleculaire, Hôpital St-Louis, 1, Avenue Claude Vellefaux, Paris, 75010, France.
authors.forename: C
authors.initials: C
authors.lastname: Menager
authors.forename: J
authors.initials: J
authors.lastname: Vassy
authors.forename: C
authors.initials: C
authors.lastname: Doliger
authors.forename: Y
authors.initials: Y
authors.lastname: Legrand
authors.forename: A
authors.initials: A
authors.lastname: Karniguian
mesh_terms: D000199:Actins / Q000378:metabolism; D001692:Biological Transport; D002448:Cell Adhesion; D002462:Cell Membrane / Q000378:metabolism; D002478:Cells, Cultured; D003094:Collagen / Q000502:physiology*; D003593:Cytoplasm / Q000378:metabolism; D003598:Cytoskeletal Proteins; D004730:Endothelium, Vascular / Q000166:cytology / Q000378:metabolism*; D016326:Extracellular Matrix Proteins / Q000378:metabolism; D005353:Fibronectins / Q000502:physiology*; D019204:GTP-Binding Proteins / Q000378:metabolism*; D006801:Humans; D046911:Macromolecular Substances; D018613:Microscopy, Confocal; D010750:Phosphoproteins / Q000378:metabolism*; D011554:Pseudopodia / Q000378:metabolism; D013347:Subcellular Fractions / Q000378:metabolism; D020744:rac GTP-Binding Proteins; D020742:rhoA GTP-Binding Protein; D000099149:Ezrin
chemical_list: D000199:Actins; D003094:Collagen; D003598:Cytoskeletal Proteins; D016326:Extracellular Matrix Proteins; D005353:Fibronectins; D019204:GTP-Binding Proteins; D046911:Macromolecular Substances; D010750:Phosphoproteins; D020744:rac GTP-Binding Proteins; D020742:rhoA GTP-Binding Protein; D000099149:Ezrin