TY - JOUR T1 - PKCδ Associates with and Is Involved in the Phosphorylation of RasGRP3 in Response to Phorbol Esters JF - Molecular Pharmacology JO - Mol Pharmacol SP - 76 LP - 84 DO - 10.1124/mol.66.1.76 VL - 66 IS - 1 AU - Chaya Brodie AU - Rivka Steinhart AU - Gila Kazimirsky AU - Hadara Rubinfeld AU - Tehila Hyman AU - Jolene N. Ayres AU - Gang Min Hur AU - Attila Toth AU - Dazhi Yang AU - Susan H. Garfield AU - James C. Stone AU - Peter M. Blumberg Y1 - 2004/07/01 UR - http://molpharm.aspetjournals.org/content/66/1/76.abstract N2 - RasGRP is a family of guanine nucleotide exchange factors that activate small GTPases and contain a C1 domain similar to the one present in protein kinase C (PKC). In this study, we examined the interaction of RasGRP3 and PKC in response to the phorbol ester PMA. In Chinese hamster ovary or LN-229 cells heterologously expressing RasGRP3, phorbol 12-myristate 13-acetate (PMA) induced translocation of RasGRP3 to the perinuclear region and a decrease in the electrophoretic mobility of RasGRP3. The mobility shift was associated with phosphorylation of RasGRP3 on serine residues and seemed to be PKCδ-dependent because it was blocked by the PKCδ inhibitor rottlerin as well as by a PKCδ kinase-dead mutant. Using coimmunoprecipitation, we found that PMA induced the physical association of RasGRP3 with PKCδ and, using in situ methods, we showed colocalization of PKCδ and RasGRP3 in the perinuclear region. PKCδ phosphorylated RasGRP3 in vitro. Previous studies suggest that ectopic expression of RasGRP3 increases activation of Erk1/2. We found that overexpression of either PKCδ or RasGRP3 increased the activation of Erk1/2 by PMA. In contrast, coexpression of PKCδ and RasGRP3 yielded a level of phosphorylation of Erk1/2 similar to that of control vector cells. Our results suggest that PKCδ may act as an upstream kinase associating with and phosphorylating RasGRP3 in response to PMA. The interaction between RasGRP3 and PKCδ points to the existence of complex cross-talk between various members of the phorbol ester receptors which can have important impact on major signal transduction pathways and cellular processes induced by phorbol esters or DAG ER -