Activation of G12/G13 results in shape change and Rho/Rho-kinase-mediated myosin light chain phosphorylation in mouse platelets.
Platelets respond to various stimuli with rapid changes in shape followed by aggregation and secretion of their granule contents. Platelets lacking the alpha-subunit of the heterotrimeric G protein Gq do not aggregate and degranulate but still undergo shape change after activation through thromboxane-A2 (TXA2) or thrombin receptors. In contrast to thrombin, the TXA2 mimetic U46619 led to the selective activation of G12 and G13 in Galphaq-deficient platelets indicating that these G proteins mediate TXA2 receptor-induced shape change. TXA2 receptor-mediated activation of G12/G13 resulted in tyrosine phosphorylation of pp72(syk) and stimulation of pp60(c-src) as well as in phosphorylation of myosin light chain (MLC) in Galphaq-deficient platelets. Both MLC phosphorylation and shape change induced through G12/G13 in the absence of Galphaq were inhibited by the C3 exoenzyme from Clostridium botulinum, by the Rho-kinase inhibitor Y-27632 and by cAMP-analogue Sp-5,6-DCl-cBIMPS. These data indicate that G12/G13 couple receptors to tyrosine kinases as well as to the Rho/Rho-kinase-mediated regulation of MLC phosphorylation. We provide evidence that G12/G13-mediated Rho/Rho-kinase-dependent regulation of MLC phosphorylation participates in receptor-induced platelet shape change.
Institut für Pharmakologie, Universitätsklinikum Benjamin Franklin, Freie Universität Berlin, 14195 Berlin, Germany.
B
B
Klages
U
U
Brandt
M I
MI
Simon
G
G
Schultz
S
S
Offermanns
D019796:15-Hydroxy-11 alpha,9 alpha-(epoxymethano)prosta-5,13-dienoic Acid / Q000494:pharmacology; D000818:Animals; D001792:Blood Platelets / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism*; D048429:Cell Size / Q000502:physiology; D044365:GTP-Binding Protein alpha Subunits, G12-G13; D043802:GTP-Binding Protein alpha Subunits, Gq-G11; D019204:GTP-Binding Proteins / Q000097:blood* / Q000172:deficiency / Q000235:genetics; D020690:GTPase-Activating Proteins; D047908:Intracellular Signaling Peptides and Proteins; D051379:Mice; D008855:Microscopy, Electron, Scanning; D018994:Myosin Light Chains / Q000097:blood*; D010766:Phosphorylation; D010974:Platelet Aggregation / Q000502:physiology; D017346:Protein Serine-Threonine Kinases / Q000097:blood; D017482:Receptors, Thromboxane / Q000378:metabolism; D013917:Thrombin / Q000494:pharmacology; D054460:rho-Associated Kinases
D020690:GTPase-Activating Proteins; D047908:Intracellular Signaling Peptides and Proteins; D018994:Myosin Light Chains; D017482:Receptors, Thromboxane; C085379:rho GTPase-activating protein; D019796:15-Hydroxy-11 alpha,9 alpha-(epoxymethano)prosta-5,13-dienoic Acid; D017346:Protein Serine-Threonine Kinases; D054460:rho-Associated Kinases; D013917:Thrombin; D019204:GTP-Binding Proteins; D044365:GTP-Binding Protein alpha Subunits, G12-G13; D043802:GTP-Binding Protein alpha Subunits, Gq-G11