J. Cell Biol., Volume 144, Number 4, February 22, 1999 745-754

Activation of G₁₂/G₁₃ Results in Shape Change and Rho/Rho-Kinase-mediated Myosin Light Chain Phosphorylation in Mouse Platelets

Birgit Klages,^* Ursula Brandt,^* Melvin I. Simon, Günter Schultz,^* and Stefan Offermanns^*

^* Institut für Pharmakologie, Universitätsklinikum Benjamin Franklin, Freie Universität Berlin, 14195 Berlin, Germany; and  Division of Biology, California Institute of Technology, Pasadena, California 91125

Platelets respond to various stimuli with rapid changes in shape followed by aggregation and secretion of their granule contents. Platelets lacking the -subunit of the heterotrimeric G protein G_q 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 G_q-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 G_q-deficient platelets. Both MLC phosphorylation and shape change induced through G₁₂/G₁₃ in the absence of G_q 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.

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