title: The related adhesion focal tyrosine kinase differentially phosphorylates p130Cas and the Cas-like protein, p105HEF1.
abstract: The related adhesion focal tyrosine kinase (RAFTK) is tyrosine-phosphorylated following beta1 integrin or B cell antigen receptor stimulation in human B cells. Two substrates that are tyrosine-phosphorylated following integrin ligation in B cells are p130(Cas) and the Cas family member human enhancer of filamentation 1 (HEF1), both of which can associate with RAFTK. In this report we observed that RAFTK was involved in the phosphorylation of these two proteins. While a catalytically active RAFTK was required for both p130(Cas) and HEF1, phosphorylation of p130(Cas), but not of HEF1, was dependent on an intact autophosphorylation site (Tyr402) on RAFTK. To determine if RAFTK phosphorylated p130(Cas) and HEF1 directly or through an intermediate, we assayed the ability of RAFTK and of a Tyr402 mutant to phosphorylate purified HEF1 and p130(Cas) domains. RAFTK was able to phosphorylate the substrate domains of both p130(Cas) and HEF1, but only the C-terminal domain of p130(Cas). Furthermore, Tyr402, which mediates the binding of RAFTK to c-Src kinase, was required for the phosphorylation of the C-terminal domain of p130(Cas). These data suggest that RAFTK itself is sufficient for HEF1 phosphorylation, whereas a cooperation between RAFTK and Src kinases is required for the complete phosphorylation of p130(Cas).
authors.affiliation: Department of Medicine, Harvard Medical School, Boston, Massachusetts 02115, USA.
authors.forename: A
authors.initials: A
authors.lastname: Astier
authors.forename: S N
authors.initials: SN
authors.lastname: Manié
authors.forename: H
authors.initials: H
authors.lastname: Avraham
authors.forename: H
authors.initials: H
authors.lastname: Hirai
authors.forename: S F
authors.initials: SF
authors.lastname: Law
authors.forename: Y
authors.initials: Y
authors.lastname: Zhang
authors.forename: E A
authors.initials: EA
authors.lastname: Golemis
authors.forename: Y
authors.initials: Y
authors.lastname: Fu
authors.forename: B J
authors.initials: BJ
authors.lastname: Druker
authors.forename: N
authors.initials: N
authors.lastname: Haghayeghi
authors.forename: A S
authors.initials: AS
authors.lastname: Freedman
authors.forename: S
authors.initials: S
authors.lastname: Avraham
mesh_terms: D048868:Adaptor Proteins, Signal Transducing; D000818:Animals; D015153:Blotting, Western; D019556:COS Cells; D000081247:CSK Tyrosine-Protein Kinase; D002384:Catalysis; D050719:Crk-Associated Substrate Protein; D004591:Electrophoresis, Polyacrylamide Gel; D051418:Focal Adhesion Kinase 2; D006801:Humans; D010750:Phosphoproteins / Q000378:metabolism*; D010766:Phosphorylation; D011505:Protein-Tyrosine Kinases / Q000378:metabolism*; D011506:Proteins; D016160:Retinoblastoma Protein / Q000378:metabolism*; D050717:Retinoblastoma-Like Protein p130; D014443:Tyrosine / Q000378:metabolism; D019061:src-Family Kinases
chemical_list: D048868:Adaptor Proteins, Signal Transducing; C493841:BCAR1 protein, human; D050719:Crk-Associated Substrate Protein; C099942:NEDD9 protein, human; D010750:Phosphoproteins; D011506:Proteins; D016160:Retinoblastoma Protein; D050717:Retinoblastoma-Like Protein p130; D014443:Tyrosine; D011505:Protein-Tyrosine Kinases; D000081247:CSK Tyrosine-Protein Kinase; D051418:Focal Adhesion Kinase 2; D019061:src-Family Kinases; C000631346:CSK protein, human