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J. Biol. Chem. 276 (23): 20130-20135

© 2001 by The American Society for Biochemistry and Molecular Biology, Inc.

Src and Pyk2 Mediate G-protein-coupled Receptor Activation of Epidermal Growth Factor Receptor (EGFR) but Are Not Required for Coupling to the Mitogen-activated Protein (MAP) Kinase Signaling Cascade*

Julian Andreev{ddagger}, Maria L. Galisteo{ddagger}, Onno Kranenburg§, Susan K. Logan{ddagger}, Ernest S. Chiu{ddagger}, Mitsuhiko Okigaki{ddagger}, Leslie A. Cary, Wouter H. Moolenaar§, , and Joseph Schlessinger{ddagger}||

From the {ddagger}Department of Pharmacology, New York University School of Medicine, Skirball Institute of Biomolecular Medicine, New York, NY 10016, the §Division of Cellular Biochemistry, The Netherlands Cancer Institute, Centre for Biomedical Genetics, Plesmanlaan 121 1066CX Amsterdam, The Netherlands, and the Fred Hutchinson Cancer Research Center, Seattle, Washington 98109

ABSTRACT Back to Top

Abstract: The epidermal growth factor receptor (EGFR) and the non-receptor protein tyrosine kinases Src and Pyk2 have been implicated in linking a variety of G-protein-coupled receptors (GPCR) to the mitogen-activated protein (MAP) kinase signaling cascade. In this report we apply a genetic strategy using cells isolated from Src-, Pyk2-, or EGFR-deficient mice to explore the roles played by these protein tyrosine kinases in GPCR-induced activation of EGFR, Pyk2, and MAP kinase. We show that Src kinases are critical for activation of Pyk2 in response to GPCR-stimulation and that Pyk2 and Src are essential for GPCR-induced tyrosine phosphorylation of EGFR. By contrast, Pyk2, Src, and EGFR are dispensable for GPCR-induced activation of MAP kinase. Moreover, GPCR-induced MAP kinase activation is normal in fibroblasts deficient in both Src and Pyk2 (Src–/–Pyk2–/– cells) as well as in fibroblasts deficient in all three Src kinases expressed in these cells (Src–/–Yes–/–Fyn–/– cells). Finally, experiments are presented demonstrating that, upon stimulation of GPCR, activated Pyk2 forms a complex with Src, which in turn phosphorylates EGFR directly. These experiments reveal a role for Src kinases in Pyk2 activation and a role for Pyk2 and Src in tyrosine phosphorylation of EGFR following GPCR stimulation. In addition, EGFR, Src family kinases, and Pyk2 are not required for linking GPCRs with the MAP kinase signaling cascade.

Received for publication March 14, 2001.

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   Abstract »    Full Text »    PDF »
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   Abstract »    Full Text »    PDF »

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