Regulation of integrin-mediated p130(Cas) tyrosine phosphorylation in human B cells. A role for p59(Fyn) and SHP2

Serge N. Manié, Anne Astier, Nilou Haghayeghi, Timothy Canty, Brian Druker, Hisamaru Hirai, Arnold S. Freedman

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Abstract

Engagement of β1 integrins in terminally differentiated human B cell lines, such as ARH-77, leads to prominent tyrosine phosphorylation of the p130 Crk-associated substrate (Cas). Cas regulates the assembly of several SH2 and SH3 domain-containing proteins into signaling complexes, which are potentially involved in the propagation of downstream signals. We demonstrate here that immunoprecipitated Cas from β1 integrin-stimulated ARH-77 cells was associated with tyrosine kinase and phosphatase activities and that integrin ligation led to the recruitment of at least p59(Fyn) tyrosine kinase and SHP2 tyrosine phosphatase in Cas immune complexes. Cotransfection studies in COS-7 cells further indicated that Fyn/Cas physical interaction and Fyn- mediated Cas phosphorylation required amino acids 638-889 in the C-terminal region of Cas. This sequence contains both c-Src SH2 and SH3 domain-binding motifs. In vitro binding studies using glutathione S-transferase fusion proteins derived from the SH2 or SH3 domains of Fyn suggested that both Fyn domains can participate in Fyn/Cas interaction. These data implicate Fyn and SHP2 as potential modulators of Cas signaling complexes in B cells.

Original languageEnglish (US)
Pages (from-to)15636-15641
Number of pages6
JournalJournal of Biological Chemistry
Volume272
Issue number25
DOIs
StatePublished - Jun 20 1997

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Crk-Associated Substrate Protein
Phosphorylation
Integrins
Tyrosine
B-Lymphocytes
src Homology Domains
Cells
Phosphoric Monoester Hydrolases
Proto-Oncogene Proteins c-fyn
Non-Receptor Type 11 Protein Tyrosine Phosphatase
COS Cells
Antigen-Antibody Complex
Glutathione Transferase
Protein-Tyrosine Kinases
Modulators
Ligation
Proteins
Fusion reactions

ASJC Scopus subject areas

  • Biochemistry

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Regulation of integrin-mediated p130(Cas) tyrosine phosphorylation in human B cells. A role for p59(Fyn) and SHP2. / Manié, Serge N.; Astier, Anne; Haghayeghi, Nilou; Canty, Timothy; Druker, Brian; Hirai, Hisamaru; Freedman, Arnold S.

In: Journal of Biological Chemistry, Vol. 272, No. 25, 20.06.1997, p. 15636-15641.

Research output: Contribution to journalArticle

Manié, Serge N. ; Astier, Anne ; Haghayeghi, Nilou ; Canty, Timothy ; Druker, Brian ; Hirai, Hisamaru ; Freedman, Arnold S. / Regulation of integrin-mediated p130(Cas) tyrosine phosphorylation in human B cells. A role for p59(Fyn) and SHP2. In: Journal of Biological Chemistry. 1997 ; Vol. 272, No. 25. pp. 15636-15641.
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AU - Astier, Anne

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AU - Canty, Timothy

AU - Druker, Brian

AU - Hirai, Hisamaru

AU - Freedman, Arnold S.

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