Src Homology 3 Binding Sites in the P2Y2 Nucleotide Receptor Interact with Src and Regulate Activities of Src, Proline-rich Tyrosine Kinase 2, and Growth Factor Receptors

Jun Liu, Zhongji Liao, Jean Camden, Korey D. Griffin, Richard C. Garrad, Laura I. Santiago-Pérez, Fernando A. González, Cheikh Seye, Gary A. Weisman, Laurie Erb

Research output: Contribution to journalArticle

117 Citations (Scopus)

Abstract

Many G protein-coupled receptors activate growth factor receptors, although the mechanisms controlling this transactivation are unclear. We have identified two proline-rich, SH3 binding sites (PXXP) in the carboxyl-terminal tail of the human P2Y2 nucleotide receptor that directly associate with the tyrosine kinase Src in protein binding assays. Furthermore, Src co-precipitated with the P2Y2 receptor in 1321N1 astrocytoma cells stimulated with the P2Y2 receptor agonist UTP. A mutant P2Y 2 receptor lacking the PXXP motifs was found to stimulate calcium mobilization and serine/threonine phosphorylation of the Erk1/2 mitogen-activated protein kinases, like the wild-type receptor, but was defective in its ability to stimulate tyrosine phosphorylation of Src and Src-dependent tyrosine phosphorylation of the proline-rich tyrosine kinase 2, epidermal growth factor receptor (EGFR), and platelet-derived growth factor receptor. Dual immunofluorescence labeling of the P2Y2 receptor and the EGFR indicated that UTP caused an increase in the co-localization of these receptors in the plasma membrane that was prevented by the Src inhibitor PP2. Together, these data suggest that agonist-induced binding of Src to the SH3 binding sites in the P2Y2 receptor facilitates Src activation, which recruits the EGFR into a protein complex with the P2Y2 receptor and allows Src to efficiently phosphorylate the EGFR.

Original languageEnglish (US)
Pages (from-to)8212-8218
Number of pages7
JournalJournal of Biological Chemistry
Volume279
Issue number9
DOIs
StatePublished - Feb 27 2004
Externally publishedYes

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Focal Adhesion Kinase 2
Purinergic P2Y2 Receptors
Growth Factor Receptors
Epidermal Growth Factor Receptor
Nucleotides
Binding Sites
Phosphorylation
Uridine Triphosphate
Tyrosine
Platelet-Derived Growth Factor Receptors
src-Family Kinases
Mitogen-Activated Protein Kinase 1
Threonine
Cell membranes
G-Protein-Coupled Receptors
Proline
Labeling
Serine
Astrocytoma
Assays

ASJC Scopus subject areas

  • Biochemistry

Cite this

Src Homology 3 Binding Sites in the P2Y2 Nucleotide Receptor Interact with Src and Regulate Activities of Src, Proline-rich Tyrosine Kinase 2, and Growth Factor Receptors. / Liu, Jun; Liao, Zhongji; Camden, Jean; Griffin, Korey D.; Garrad, Richard C.; Santiago-Pérez, Laura I.; González, Fernando A.; Seye, Cheikh; Weisman, Gary A.; Erb, Laurie.

In: Journal of Biological Chemistry, Vol. 279, No. 9, 27.02.2004, p. 8212-8218.

Research output: Contribution to journalArticle

Liu, Jun ; Liao, Zhongji ; Camden, Jean ; Griffin, Korey D. ; Garrad, Richard C. ; Santiago-Pérez, Laura I. ; González, Fernando A. ; Seye, Cheikh ; Weisman, Gary A. ; Erb, Laurie. / Src Homology 3 Binding Sites in the P2Y2 Nucleotide Receptor Interact with Src and Regulate Activities of Src, Proline-rich Tyrosine Kinase 2, and Growth Factor Receptors. In: Journal of Biological Chemistry. 2004 ; Vol. 279, No. 9. pp. 8212-8218.
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AU - Liu, Jun

AU - Liao, Zhongji

AU - Camden, Jean

AU - Griffin, Korey D.

AU - Garrad, Richard C.

AU - Santiago-Pérez, Laura I.

AU - González, Fernando A.

AU - Seye, Cheikh

AU - Weisman, Gary A.

AU - Erb, Laurie

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