title: Reactive oxygen species mediate Rac-induced loss of cell-cell adhesion in primary human endothelial cells.
abstract: The integrity of the endothelium is dependent on cell-cell adhesion, which is mediated by vascular-endothelial (VE)-cadherin. Proper VE-cadherin-mediated homotypic adhesion is, in turn, dependent on the connection between VE-cadherin and the cortical actin cytoskeleton. Rho-like small GTPases are key molecular switches that control cytoskeletal dynamics and cadherin function in epithelial as well as endothelial cells. We show here that a cell-penetrating, constitutively active form of Rac (Tat-RacV12) induces a rapid loss of VE-cadherin-mediated cell-cell adhesion in endothelial cells from primary human umbilical veins (pHUVEC). This effect is accompanied by the formation of actin stress fibers and is dependent on Rho activity. However, transduction of pHUVEC with Tat-RhoV14, which induces pronounced stress fiber and focal adhesion formation, did not result in a redistribution of VE-cadherin or an overall loss of cell-cell adhesion. In line with this observation, endothelial permeability was more efficiently increased by Tat-RacV12 than by Tat-RhoV14. The loss of cell-cell adhesion, which is induced by Tat-RacV12, occurred in parallel to and was dependent upon the intracellular production of reactive oxygen species (ROS). Moreover, Tat-RacV12 induced an increase in tyrosine phosphorylation of a component the VE-cadherin-catenin complex, which was identified as alpha-catenin. The functional relevance of this signaling pathway was further underscored by the observation that endothelial cell migration, which requires a transient reduction of cell-cell adhesion, was blocked when signaling through ROS was inhibited. In conclusion, Rac-mediated production of ROS represents a previously unrecognized means of regulating VE-cadherin function and may play an important role in the (patho)physiology associated with inflammation and endothelial damage as well as with endothelial cell migration and angiogenesis.
authors.affiliation: Department of Experimental Immunohematology, Academic Medical Center, Plesmanlaan 125, 1066 CX Amsterdam, The Netherlands.
authors.forename: Sandra
authors.initials: S
authors.lastname: van Wetering
authors.forename: Jaap D
authors.initials: JD
authors.lastname: van Buul
authors.forename: Safira
authors.initials: S
authors.lastname: Quik
authors.forename: Frederik P J
authors.initials: FP
authors.lastname: Mul
authors.forename: Eloise C
authors.initials: EC
authors.lastname: Anthony
authors.forename: Jean-Paul
authors.initials: JP
authors.lastname: ten Klooster
authors.forename: John G
authors.initials: JG
authors.lastname: Collard
authors.forename: Peter L
authors.initials: PL
authors.lastname: Hordijk
mesh_terms: D008841:Actin Cytoskeleton / Q000378:metabolism*; D015703:Antigens, CD; D015820:Cadherins / Q000378:metabolism*; D002448:Cell Adhesion / Q000502:physiology*; D002478:Cells, Cultured; D004730:Endothelium, Vascular / Q000166:cytology / Q000378:metabolism*; D005347:Fibroblasts / Q000166:cytology / Q000378:metabolism; D006801:Humans; D010766:Phosphorylation; D017382:Reactive Oxygen Species / Q000378:metabolism*; D015398:Signal Transduction / Q000502:physiology; D014161:Transduction, Genetic; D014443:Tyrosine / Q000378:metabolism; D020744:rac GTP-Binding Proteins / Q000378:metabolism*; D020741:rho GTP-Binding Proteins / Q000378:metabolism; D000099152:Cadherin 5
chemical_list: D015703:Antigens, CD; D015820:Cadherins; D017382:Reactive Oxygen Species; D014443:Tyrosine; D020744:rac GTP-Binding Proteins; D020741:rho GTP-Binding Proteins; D000099152:Cadherin 5