Proline-rich sequences mediate the interaction of the Arg protein tyrosine kinase with Crk

Baolin Wang, Tami H. Mysliwiec, Stephan M. Feller, Beatrice Knudsen, Hidesaburo Hanafusa, Gary D. Kruh

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Abstract

Arg is a ubiquitously expressed member of the Abelson family of nonreceptor protein-tyrosine kinases. Defining the Arg sequences that mediate its interaction with other proteins is essential to elucidating its role in cellular signaling. In this report we demonstrate that Arg associates with c-Crk, an adaptor protein composed of an SH2 domain and two SH3 domains, and examine the molecular mechanism of the interaction. In vitro experiments revealed that three proline-rich sequences with distinct specificities for SH3 domains are located in the Arg C-terminal domain, just C-terminal to the kinase domain, and that two of these sequences bind to the Crk N-terminal SH3 domain. These two sequences conform to the PxLPxK/R motif that has been observed in other proteins that bind the Crk N-terminal SH3 domain. The interaction of Arg with c-Crk in living cells was confirmed by the detection of coimmunoprecipitation in coinfected Sf9 cells. In addition, increased phosphorylation of c-Crk was observed in cotransfected COS cells, indicating that Crk is an Arg substrate. The site of c-Crk phosphorylation by Arg was identified as tyrosine 221, a residue whose modification has been shown to result in an intramolecular SH2 interaction and a folded conformation. These experiments extend the known Arg protein interacting motifs to include SH3 binding sites and suggest that Arg may function as an effector as well as a regulator of Crk activity.

Original languageEnglish (US)
Pages (from-to)1379-1385
Number of pages7
JournalOncogene
Volume13
Issue number7
Publication statusPublished - Nov 2 1996

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All Science Journal Classification (ASJC) codes

  • Molecular Biology
  • Genetics
  • Cancer Research

Cite this

Wang, B., Mysliwiec, T. H., Feller, S. M., Knudsen, B., Hanafusa, H., & Kruh, G. D. (1996). Proline-rich sequences mediate the interaction of the Arg protein tyrosine kinase with Crk. Oncogene, 13(7), 1379-1385.