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Blood, 15 December 2002, Vol. 100, No. 13, pp. 4462-4469
HEMOSTASIS, THROMBOSIS, AND VASCULAR BIOLOGY
Cdc42/Rac1-dependent activation of the p21-activated kinase (PAK)
regulates human platelet lamellipodia spreading: implication of the
cortical-actin binding protein cortactin
Catherine Vidal,
Blandine Geny,
Josiane Melle,
Martine Jandrot-Perrus, and
Michaëla Fontenay-Roupie
From the Département d'Hématologie and
Département de Biologie Cellulaire, Institut Cochin, Institut
National de la Santé et de la Recherche Médicale (INSERM),
Centre National de la Recherche Scientifique (CNRS), Université
René Descartes, Laboratoire Central d'Hématologie,
Hôpital Cochin, AP-HP and E9907 INSERM, Faculté Xavier
Bichat, Paris, France.
Platelet activation by thrombin or thrombin receptor-activating
peptide (TRAP) results in extensive actin reorganization that leads to
filopodia emission and lamellae spreading concomitantly with activation
of the Rho family small G proteins, Cdc42 and Rac1. Evidence has been
provided that direct binding of Cdc42-guanosine triphosphate
(GTP) and Rac1-GTP to the N-terminal regulatory domain of the
p21-activated kinase (PAK) stimulates PAK activation and actin
reorganization. In the present study, we have investigated the
relationship between shape change and PAK activation. We show that
thrombin, TRAP, or monoclonal antibody (MoAb) anti-Fc RIIA IV.3 induces an activation of Cdc42 and Rac1. The GpVI ligand, convulxin (CVX), that forces platelets to lamellae spreading
efficiently activates Rac1. Thrombin, TRAP, MoAb IV.3, and CVX
stimulate autophosphorylation and kinase activity of PAK. Inhibition of
Cdc42 and Rac1 with clostridial toxin B inhibits PAK activation and
lamellae spreading. The cortical-actin binding protein, p80/85
cortactin, is constitutively associated with PAK in resting platelets
and dissociates from PAK after thrombin stimulation. Inhibition of PAK
autophosphorylation by toxin B prevents the dissociation of
cortactin. These results suggest that Cdc42/Rac1-dependent
activation of PAK may trigger early platelet shape change, at
least in part through the regulation of cortactin binding to PAK.
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