Recruitment of Pyk2 to SHPS-1 signaling complex is required for IGF-I-dependent mitogenic signaling in vascular smooth muscle cells
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In vascular smooth muscle cells, IGF-I stimulates SHPS-1/SHP2/Src complex formation which is required for IGF-I-stimulated cell proliferation. Using SHP2/Src silencing and a Pyk2/Y402F mutant, we showed that Pyk2 was also recruited to the SHPS-1 complex. Pyk2 recruitment to SHPS-1 is mediated via the interaction of Pyk2 Tyr402 and the Src in response to IGF-I. Following Src/Pyk2 association, Src phosphorylates Pyk2 on Tyr881 providing a binding site for Grb2. Cells expressing Pyk2/Y881F showed decreased Grb2 recruitment to SHPS-1 and impaired Shc/Grb2 association. This change led to reduced Erk1/2 (MAP kinase) activation and cell proliferation in response to IGF-I. Our results show that, following its recruitment to the SHPS-1 signaling complex, Pyk2 localizes Grb2 in close proximity to Shc thereby facilitating Shc/Grb2 association which leads to Erk1/2 activation in response to IGF-I. Thus, Pyk2 recruitment to SHPS-1 plays an important role in regulating the IGF-I-stimulated mitogenic response.
KeywordsIGF-I SHPS-1 Pyk2 Grb2 Shc MAP kinase Cell proliferation
We thank Dr. Walker H. Busby, Jr. for his help in preparing the SHPS-1 antibody. We thank Drs. H.S. Earp and Lee M Graves (University of North Carolina at Chapel Hill) for providing pcDNA-myc-Pyk2/WT, pcDNA-myc-Pyk2/Y402F, and pcDNA-myc-Pyk2/Y881F constructs. We thank Drs. Laura A. Maile and Lee M Graves for comments. We also thank Ms. Laura Lindsey for her help in preparing the manuscript. This study was supported by a grant HL56850 and AG022331 from the National Institutes of Health.
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