1. ^* Department of Pharmacology, New York University Medical Center, 550 First Avenue, New York, New York 10016, USA
2. ^† SUGEN Inc., 515 Galveston Drive, Redwood City, California 94063, USA

Abstract

THE mechanisms by which mitogenic G-protein-coupled receptors activate the MAP kinase signalling pathway are poorly understood. Candidate protein tyrosine kinases that link G-protein-coupled receptors with MAP kinase include Src family kinases¹, the epidermal growth factor receptor², Lyn and Syk³. Here we show that lysophosphatidic acid (LPA) and bradykinin induce tyrosine phosphorylation of Pyk2 and complex formation between Pyk2 and activated Src. Moreover, tyrosine phosphorylation of Pyk2 leads to binding of the SH2 domain of Src to tyrosine 402 of Pyk2 and activation of Src. Transient overexpres-sion of a dominant interfering mutant of Pyk2 or the protein tyrosine kinase Csk reduces LPA- or bradykinin-induced activation of MAP kinase. LPA- or bradykinin-induced MAP kinase activation was also inhibited by overexpression of dominant interfering mutants of Grb2 and Sos. We propose that Pyk2 acts with Src to link G_i- and G_q-coupled receptors with Grb2 and Sos to activate the MAP kinase signalling pathway in PC12 cells.