Differential Interaction of CrkII Adaptor Protein with Platelet-Derived Growth Factor α- and β-Receptors Is Determined by Its Internal Tyrosine Phosphorylation

Abstract CrkII is an intracellular adaptor protein involved in signal transduction by various growth factors. Activation of PDGF α-receptor resulted in its association with CrkII in vivo. In contrast, binding of CrkII to the PDGF β-receptor was negligible, despite its becoming prominently phosphorylated. Bacterially expressed GST-CrkII SH2 domain specifically bound to Tyr-762 and Tyr-771 in the activated PDGF α- and β- receptors, respectively. GST fusion protein of full-length CrkII also bound to the activated PDGF β-receptor. However, tyrosine phosphorylation of GST-CrkII diminished its binding to the β-receptor. CrkI, a truncated version of CrkII lacking the phosphorylatable tyrosine residue, could bind to both PDGF α- and β-receptors in vivo. In conclusion, tyrosine phosphorylation of CrkII negatively affects its binding to the PDGF receptors. The differential binding of CrkII to the PDGF α- and β- receptors may be a rationale for functional diversity between the two receptors.

[1]  M. Matsuda,et al.  Phosphorylation of CrkII Adaptor Protein at Tyrosine 221 by Epidermal Growth Factor Receptor* , 1998, The Journal of Biological Chemistry.

[2]  Yasushi Saito,et al.  Identification of Tyr-762 in the platelet-derived growth factor α-receptor as the binding site for Crk proteins , 1998, Oncogene.

[3]  S. Spiegel,et al.  Sphingosine 1-Phosphate Stimulates Tyrosine Phosphorylation of Crk* , 1997, The Journal of Biological Chemistry.

[4]  C. Heldin.  Simultaneous induction of stimulatory and inhibitory signals by PDGF , 1997, FEBS letters.

[5]  M. Matsuda,et al.  Emerging components of the Crk oncogene product: the first identified adaptor protein. , 1996, Cellular signalling.

[6]  M. Shibuya,et al.  DOCK180, a major CRK-binding protein, alters cell morphology upon translocation to the cell membrane , 1996, Molecular and cellular biology.

[7]  L. Claesson-Welsh,et al.  Platelet-derived growth factor receptor signals. , 1994, The Journal of biological chemistry.

[8]  P. Tempst,et al.  Identification of CRKL as the constitutively phosphorylated 39-kD tyrosine phosphoprotein in chronic myelogenous leukemia cells. , 1994, Blood.

[9]  C. Heldin,et al.  Tyr-716 in the platelet-derived growth factor beta-receptor kinase insert is involved in GRB2 binding and Ras activation , 1994, Molecular and cellular biology.

[10]  Michael Karin,et al.  Membrane targeting of the nucleotide exchange factor Sos is sufficient for activating the Ras signaling pathway , 1994, Cell.

[11]  B. Druker,et al.  Crkl is the major tyrosine-phosphorylated protein in neutrophils from patients with chronic myelogenous leukemia. , 1994, The Journal of biological chemistry.

[12]  N. Heisterkamp,et al.  Tyrosine phosphorylation of CRKL in Philadelphia+ leukemia. , 1994, Blood.

[13]  M. Matsuda,et al.  CRK protein binds to two guanine nucleotide-releasing proteins for the Ras family and modulates nerve growth factor-induced activation of Ras in PC12 cells , 1994, Molecular and cellular biology.

[14]  S. Feller,et al.  c‐Abl kinase regulates the protein binding activity of c‐Crk. , 1994, The EMBO journal.

[15]  C. Heldin,et al.  A tyrosine residue in the juxtamembrane segment of the platelet-derived growth factor beta-receptor is critical for ligand-mediated endocytosis. , 1994, The Journal of biological chemistry.

[16]  A. Kazlauskas,et al.  The PDGF receptor alpha subunit activates p21ras and triggers DNA synthesis without interacting with rasGAP. , 1994, Oncogene.

[17]  N. Heisterkamp,et al.  Isolation and chromosomal localization of CRKL, a human crk-like gene. , 1993, Oncogene.

[18]  M. Matsuda,et al.  Both the SH2 and SH3 domains of human CRK protein are required for neuronal differentiation of PC12 cells , 1993, Molecular and cellular biology.

[19]  S. Aaronson,et al.  Differences in substrate specificities of alpha and beta platelet-derived growth factor (PDGF) receptors. Correlation with their ability to mediate PDGF transforming functions. , 1993, The Journal of biological chemistry.

[20]  T. Pawson,et al.  SH2 domains recognize specific phosphopeptide sequences , 1993, Cell.

[21]  B. Mayer,et al.  The product of the cellular crk gene consists primarily of SH2 and SH3 regions. , 1992, Cell growth & differentiation : the molecular biology journal of the American Association for Cancer Research.

[22]  C. Heldin,et al.  PDGF alpha‐ and beta‐receptors activate unique and common signal transduction pathways. , 1992, The EMBO journal.

[23]  Jonathan A. Cooper,et al.  Binding of GAP to activated PDGF receptors. , 1990, Science.