Cdc42/Rac1-dependent activation of the p21-activated kinase (PAK) regulates human platelet lamellipodia spreading: implication of the cortical-actin binding protein cortactin

被引:129
作者
Vidal, C
Geny, B
Melle, J
Jandrot-Perrus, M
Fontenay-Roupie, M
机构
[1] Univ Paris 05, Hop Cochin, APHP,Lab Cent Hematol, Dept Hematol, Paris, France
[2] Univ Paris 05, Hop Cochin,APHP,Lab Cent Hematol, INSERM,CNRS,Inst Cochin, Dept Biol Cellulaire, Paris, France
关键词
D O I
10.1182/blood.V100.13.4462
中图分类号
R5 [内科学];
学科分类号
1002 ; 100201 ;
摘要
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-FcgammaRIIA 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.
引用
收藏
页码:4462 / 4469
页数:8
相关论文
共 41 条
[1]   THE FC RECEPTOR FOR IMMUNOGLOBULIN-G (FC-GAMMA-RII) ON HUMAN PLATELETS [J].
ANDERSON, CL ;
CHACKO, GW ;
OSBORNE, JM ;
BRANDT, JT .
SEMINARS IN THROMBOSIS AND HEMOSTASIS, 1995, 21 (01) :1-9
[2]   Activation of the small GTPases, rac and cdc42, after ligation of the platelet PAR-1 receptor [J].
Azim, AC ;
Barkalow, K ;
Chou, J ;
Hartwig, JH .
BLOOD, 2000, 95 (03) :959-964
[3]   Characterization of Rac and Cdc42 activation in chemoattractant-stimulated human neutrophils using a novel assay for active GTPases [J].
Benard, V ;
Bohl, BP ;
Bokoch, GM .
JOURNAL OF BIOLOGICAL CHEMISTRY, 1999, 274 (19) :13198-13204
[4]   KINETICS OF THE INTERACTION OF MYOSIN SUBFRAGMENT-L WITH G-ACTIN [J].
BLANCHOIN, L ;
FIEVEZ, S ;
TRAVERS, F ;
CARLIER, MF ;
PANTALONI, D .
JOURNAL OF BIOLOGICAL CHEMISTRY, 1995, 270 (13) :7125-7133
[5]   The mechanism of PAK activation - Autophosphorylation events in both regulatory and kinase domains control activity [J].
Chong, C ;
Tan, L ;
Lim, L ;
Manser, E .
JOURNAL OF BIOLOGICAL CHEMISTRY, 2001, 276 (20) :17347-17353
[6]   Requirements for both Rac1 and Cdc42 in membrane ruffling and phagocytosis in leukocytes [J].
Cox, D ;
Chang, P ;
Zhang, Q ;
Reddy, PG ;
Bokoch, GM ;
Greenberg, S .
JOURNAL OF EXPERIMENTAL MEDICINE, 1997, 186 (09) :1487-1494
[7]   Membrane targeting of p21-activated kinase 1 (PAK1) induces neurite outgrowth from PC12 cells [J].
Daniels, RH ;
Hall, PS ;
Bokoch, GM .
EMBO JOURNAL, 1998, 17 (03) :754-764
[8]   Adhesion to the extracellular matrix regulates the coupling of the small GTPase Rac to its effector PAK [J].
del Pozo, MA ;
Price, LS ;
Alderson, NB ;
Ren, XD ;
Schwartz, MA .
EMBO JOURNAL, 2000, 19 (09) :2008-2014
[9]   Activation of LIM-kinase by Pak1 couples Rac/Cdc42 GTPase signalling to actin cytoskeletal dynamics [J].
Edwards, DC ;
Sanders, LC ;
Bokoch, GM ;
Gill, GN .
NATURE CELL BIOLOGY, 1999, 1 (05) :253-259
[10]   Phosphorylation and reorganization of vimentin by p21-activated kinase (PAK) [J].
Goto, H ;
Tanabe, K ;
Manser, E ;
Lim, L ;
Yasui, Y ;
Inagaki, M .
GENES TO CELLS, 2002, 7 (02) :91-97