Protein Kinase C-related Kinase and ROCK Are Required for Thrombin-induced Endothelial Cell Permeability Downstream from G{alpha}12/13 and G{alpha}11/q.

Julie Gavard (1) , J Silvio Gutkind (2)

1 UMR_S567 / UMR 8104 - Institut Cochin
2 Department of Health and Human Services

Increase in vascular permeability occurs under many physiological conditions and is central in diverse human pathologies. Thrombin is a pro-coagulant serine protease, which causes the local loss of endothelial barrier integrity thereby enabling the rapid extravasation of plasma proteins and the local formation of fibrin-containing clots. Available information suggests that thrombin induces endothelial permeability by promoting actomyosin contractility through the Rho/ROCK signaling pathway. Here we took advantage of pharmacological inhibitors, knockdown approaches, and the emerging knowledge on how permeability factors affect endothelial junctions to investigate in detail the mechanism underlying thrombin-induced endothelial permeability. We show that thrombin signals through PAR-1 and its coupled G proteins G(12/13) and G(11/q) to induce RhoA activation and intracellular calcium elevation, and that these events are interrelated. In turn, this leads to the stimulation of ROCK, which causes actin stress-fiber formation. However, this alone is not sufficient to account for thrombin-induced permeability. Instead, we found that protein kinase C-related kinase, a Rho-dependent S/T kinase, is activated in endothelial cells upon thrombin stimulation and that its expression is required for endothelial permeability and remodeling of cell-extracellular matrix and cell-cell adhesions. Our results demonstrate that the signal initiated by thrombin bifurcates at the level of RhoA to promote changes in the cytoskeletal architecture through ROCK and the remodeling of focal adhesion components through PRK. Ultimately, both pathways converge to cause cell-cell junction disruption and provoke vascular leakage.

Domaines: Interactions cellulaires [q-bio.CB]

Julie Gavard, J Silvio Gutkind. Protein Kinase C-related Kinase and ROCK Are Required for Thrombin-induced Endothelial Cell Permeability Downstream from G{alpha}12/13 and G{alpha}11/q.. Journal of Biological Chemistry, 2008, 283 (44), pp.29888-96. ⟨10.1074/jbc.M803880200⟩. ⟨hal-00340312⟩