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Activation of PAK1 in response to stimulation with carbachol and EGF and the overexpression of G protein betagamma subunits

, : Activation of PAK1 in response to stimulation with carbachol and EGF and the overexpression of G protein betagamma subunits. FASEB Journal 16(5): A1197, March 22

PAKs (p21-Activated Kinases) were the first identified mammalian members of a growing family of Ste20-like serine-threonine protein kinases. While PAKs are involved in the regulation of many processes within the cell, little is known of the signaling components immediately upstream or downstream of these proteins in mammalian cells. In this study we compared treatment of COS7 cells with carbachol and epidermal growth factor (EGF) for their effect on transiently-transfected mycPAK activity. The COS7 cells were also co-transfected with muscarinic receptors and wild type, constitutively-active and dominant-negative forms of Ras, CDC42, and Rac1, as well as Gbetagamma subunits. The stimulation of PAK can be demonstrated in an in vitro kinase assay in which recombinant Rac1 and GTPgammaS were added to the immunoprecipitated PAK and myelin basic protein (MBP) substrate. Carbachol and EGF activated mycPAK with distinct kinetics, but both were inhibited by dominant negative forms of Rac1 or CDC42. Cells co-transfected with mycPAK plus active Rac1, or active CDC42, or Gbetagamma showed increased levels of mycPAK activity, in both the presence and absence of carbachol and EGF.


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