title: Phosphorylation of Crk on tyrosine 251 in the RT loop of the SH3C domain promotes Abl kinase transactivation.
abstract: Here, we report the identification and characterization of a novel tyrosine phosphorylation site in the carboxy-terminal Src Homology 3 (SH3) (SH3C) domain of the Crk adaptor protein. Y251 is located in the highly conserved RT loop structure of the SH3C, a region of Crk involved in the allosteric regulation of the Abl kinase. Exploiting kinase assays to show that Y251 is phosphorylated by Abl in vitro, we generated affinity-purified antisera against phosphorylated Y251 in Crk and showed that Abl induces phosphorylation at Y251 in vivo, and that the kinetics of phosphorylation at Y251 and the negative regulatory Y221 site in vitro are similar. Y251 on endogenous Crk was robustly phosphorylated in chronic myelogenous leukemia cell lines and in A431 and MDA-MB-468 cells stimulated with epidermal growth factor. Using streptavidin-biotin pull downs and unbiased high-throughput Src Homology 2 (SH2) profiling approaches, we found that a pY251 phosphopeptide binds specifically to a subset of SH2 domains, including Abl and Arg SH2, and that binding of pY251 to Abl SH2 induces transactivation of Abl 1b. Finally, the Y251F Crk mutant significantly abrogates Abl transactivation in vitro and in vivo. These studies point to a yet unrealized positive regulatory role resulting from tyrosine phosphorylation of Crk, and identify a novel mechanism by which an adaptor protein activates a non-receptor tyrosine kinase by SH2 domain displacement.
authors: Department of Biochemistry and Molecular Biology, UMDNJ-New Jersey Medical School, Newark, NJ 07103-6399, USA.
authors: G
authors: G
authors: Sriram
authors: C
authors: C
authors: Reichman
authors: A
authors: A
authors: Tunceroglu
authors: N
authors: N
authors: Kaushal
authors: T
authors: T
authors: Saleh
authors: K
authors: K
authors: Machida
authors: B
authors: B
authors: Mayer
authors: Q
authors: Q
authors: Ge
authors: J
authors: J
authors: Li
authors: P
authors: P
authors: Hornbeck
authors: C G
authors: CG
authors: Kalodimos
authors: R B
authors: RB
authors: Birge
references: Mol Cell Biol. 2003 Jun;23(11):3884-96
references: 12748290
references: Biochem Biophys Res Commun. 2007 Nov 3;362(4):976-81
references: 17825249
references: Nature. 2006 Jan 12;439(7073):168-74
references: 16273093
references: FEBS Lett. 2010 Aug 4;584(15):3279-86
references: 20598684
references: J Biol Chem. 1998 Jul 3;273(27):17186-91
references: 9642287
references: Mol Cell Biol. 1995 Dec;15(12):6746-53
references: 8524240
references: Mol Cell Biol. 1993 Aug;13(8):4648-56
references: 7687742
references: J Cell Biol. 1998 Feb 23;140(4):961-72
references: 9472046
references: Biochemistry. 2006 Jul 25;45(29):8874-84
references: 16846230
references: Mol Cell. 2007 Jun 22;26(6):899-915
references: 17588523
references: Oncogene. 1998 May 14;16(19):2425-34
references: 9627109
references: Science. 1990 Jun 22;248(4962):1537-9
references: 1694307
references: J Biol Chem. 1994 Dec 30;269(52):32781-7
references: 7806500
references: Proc Natl Acad Sci U S A. 1994 Apr 12;91(8):3443-7
references: 7512734
references: J Biol Chem. 1996 Aug 30;271(35):21365-74
references: 8702917
references: Am J Pathol. 2010 Jan;176(1):446-60
references: 20008144
references: Oncogene. 2001 Feb 22;20(8):951-61
references: 11314030
references: Genes Dev. 1994 Apr 1;8(7):783-95
references: 7926767
references: Genes Dev. 1999 Sep 15;13(18):2400-11
references: 10500097
references: Mol Cell Biol. 1992 Aug;12(8):3482-9
references: 1630456
references: Cell. 2003 Mar 21;112(6):845-57
references: 12654250
references: Oncogene. 2008 Feb 14;27(8):1095-105
references: 17700528
references: Nature. 1995 Mar 30;374(6521):477-9
references: 7700361
references: Nat Chem Biol. 2011 Jan;7(1):51-7
references: 21131971
references: PLoS One. 2009 Oct 13;4(10):e7439
references: 19823681
references: Mol Cancer Res. 2005 Apr;3(4):183-94
references: 15831672
references: Nat Struct Mol Biol. 2007 Jun;14(6):503-10
references: 17515907
references: J Biol Chem. 2000 Aug 11;275(32):24787-97
references: 10825157
references: J Biol Chem. 1996 Mar 29;271(13):7375-80
references: 8631760
references: Oncogene. 2001 Oct 1;20(44):6348-71
references: 11607838
references: Mol Cell Biol. 2006 Aug;26(16):6272-82
references: 16880535
references: Nat Genet. 2001 Mar;27(3):293-8
references: 11242111
references: J Biol Chem. 1998 Sep 18;273(38):24479-84
references: 9733740
references: Oncogene. 2005 Dec 8;24(55):8187-99
references: 16158059
references: Nat Cell Biol. 2006 Aug;8(8):815-25
references: 16862147
references: Mol Cell. 2007 Feb 9;25(3):413-26
references: 17289588
references: Biochem Biophys Res Commun. 2000 Apr 2;270(1):28-33
references: 10733900
references: Cell. 1993 Mar 12;72(5):767-78
references: 7680959
references: Mol Cell Biol. 1994 Aug;14(8):5495-500
references: 8035825
references: Genes Cells. 2001 May;6(5):431-40
references: 11380621
references: Biochim Biophys Acta. 2008 May;1783(5):737-47
references: 18328268
references: Cancer Lett. 2002 Jun 6;180(1):55-61
references: 11911970
references: Nature. 1988 Mar 17;332(6161):272-5
references: 2450282
references: EMBO J. 1994 May 15;13(10):2341-51
references: 8194526
references: Oncogene. 2003 Sep 11;22(39):7950-7
references: 12970743
references: Oncogene. 2006 Jun 15;25(25):3547-56
references: 16491127
references: EMBO J. 2002 Sep 2;21(17):4571-82
references: 12198159
references: Cell Growth Differ. 1992 Jul;3(7):451-60
references: 1329926
references: Mol Biol Cell. 2002 Dec;13(12):4231-42
references: 12475948
references: Cancer Res. 2006 Jun 1;66(11):5648-55
references: 16740702
references: Cell Commun Signal. 2009 May 10;7:13
references: 19426560