c-Abl kinase regulates the protein binding activity of c-Crk.

Abstract

c‐Crk is a proto‐oncogene product composed largely of Src homology (SH) 2 and 3 domains. We have identified a kinase activity, which binds to the first Crk SH3 domain and phosphorylates c‐Crk on tyrosine 221 (Y221), as c‐Abl. c‐Abl has a strong preference for c‐Crk, when compared with common tyrosine kinase substrates. The phosphorylation of c‐Crk Y221 creates a binding site for the Crk SH2 domain. Bacterially expressed c‐Crk protein lacks phosphorylation on Y221 and can bind specifically to several proteins, while mammalian c‐Crk, which is phosphorylated on tyrosine, remains uncomplexed. The protein binding activity of c‐Crk is therefore likely regulated by a mechanism similar to that of the Src family kinases. v‐Crk is truncated before c‐Crk Y221 and forms constitutive complexes with c‐Abl and other proteins. Our results suggest that c‐Abl regulates c‐Crk function and that it could be involved in v‐Crk transformation.

The SH2-containing adapter protein GRB10 interacts with BCR-ABL

“…The interaction of Grb10 with Bcr-Abl in yeast was comparable in strength both by histidine autotrophy (Figure 2a) and b-galactosidase activity (data not shown). As expected, the Crk-SH3 domain which is known to bind to a proline rich sequence in the Abl C-terminus (Feller et al, 1994a;Ren et al, 1994), still showed interaction with the kinase-defective Bcr-Abl in yeast ( Figure 2a). Thus, binding of Grb10 to Bcr-Abl is mediated by an interaction between the Grb10-SH2 domain and a BcrAbl autophosphorylation site.…”

Development of highly selective SH3 binding peptides for Crk and CRKL which disrupt Crk-complexes with DOCK180, SoS and C3G

“…One member of this protein family, c-Crk II, is composed of an N-terminal SH2 domain, followed by two SH3 domains (Reichman et al, 1992;Matsuda et al, 1992). The SH3 domains are separated by a`spacer region' which contains a protein conformation-regulating tyrosine residue (Feller et al, 1994a;Rosen et al, 1995). c-Crk II and Crk-like (CRKL, a homologous but distinct gene product abundant in some hematopoietic cell lines) are substrates for tyrosine kinases, including those of the Abl family (c-Abl, Bcr-Abl and Arg) (Ren et al, 1994;Feller et al, 1994a;ten Hoeve et al, 1994;Wang et al, 1996b).…”

“…The SH3 domains are separated by a`spacer region' which contains a protein conformation-regulating tyrosine residue (Feller et al, 1994a;Rosen et al, 1995). c-Crk II and Crk-like (CRKL, a homologous but distinct gene product abundant in some hematopoietic cell lines) are substrates for tyrosine kinases, including those of the Abl family (c-Abl, Bcr-Abl and Arg) (Ren et al, 1994;Feller et al, 1994a;ten Hoeve et al, 1994;Wang et al, 1996b). Although detailed ultrastructural data are missing at this point, indirect evidence suggests that a CrkSH2-Y221 interaction somehow inhibits binding of the ®rst CrkSH3 domain to other cellular signalling proteins (Feller et al, 1994a).…”

“…c-Crk II and Crk-like (CRKL, a homologous but distinct gene product abundant in some hematopoietic cell lines) are substrates for tyrosine kinases, including those of the Abl family (c-Abl, Bcr-Abl and Arg) (Ren et al, 1994;Feller et al, 1994a;ten Hoeve et al, 1994;Wang et al, 1996b). Although detailed ultrastructural data are missing at this point, indirect evidence suggests that a CrkSH2-Y221 interaction somehow inhibits binding of the ®rst CrkSH3 domain to other cellular signalling proteins (Feller et al, 1994a). The functional consequences of CRKL phosphorylation at a homologous site (Y207) in Bcr ± Abl-positive hematopoietic cells (de Jong et al, 1997) are currently not understood.…”

Tyrosine-614, the major autophosphorylation site of the receptor tyrosine kinase HEK2, functions as multi-docking site for SH2-domain mediated interactions

“…The cDNA encoding the SH2-domain of human Arg (aa 163 ± 274) was ampli®ed by PCR and inserted into the vector pGEX-2T. SH2 fusion proteins of Crk, Crkl, Nck and Fer have been described previously (Feller et al, 1994(Feller et al, , 1995. The following pGEX-vectors were generous gifts: Vav/SH2 (S Katzav), Stat1/SH2 (JE Darnell Jr.), Syp/SH2 (N+C) (T Tauchi and H Broxmeyer), Tensin/SH2 (LB Chen), Fyn/SH2 (C Rudd).…”