v-Crk activates the phosphoinositide 3-kinase/AKT pathway in transformation.

v-Crk induces cellular tyrosine phosphorylation and transformation of chicken embryo fibroblasts (CEF). We studied the molecular mechanism of the v-Crk-induced transformation. Experiments with Src homology (SH)2 and SH3 domain mutants revealed that the induction of tyrosine phosphorylation of cellular proteins requires only the SH2 domain, but both the SH2 and SH3 domains are required for complete transformation. Analysis of three well defined signaling pathways, the mitogen-activated protein kinase (MAPK) pathway, the Jun N-terminal kinase (JNK) pathway, and the phosphoinositide 3-kinase (PI3K)/AKT pathway, demonstrated that only the PI3K/AKT pathway is constitutively activated in v-Crk-transformed CEF. Both the SH2 and SH3 domains are required for this activation of the PI3K/AKT pathway in CEF. We also found that the colony formation of CEF is strongly induced by a constitutively active PI3K mutant, and that a PI3K inhibitor, LY294002, suppresses the v-Crk-induced transformation. These results strongly suggest that constitutive activation of the PI3K/AKT pathway plays an essential role in v-Crk-induced transformation of CEF.

[1]  P. Vogt,et al.  The Catalytic Subunit of Phosphoinositide 3-Kinase: Requirements for Oncogenicity* , 2000, The Journal of Biological Chemistry.

[2]  S. R. Datta,et al.  Cellular survival: a play in three Akts. , 1999, Genes & development.

[3]  G. Martin,et al.  Transformation by v-Src: Ras-MAPK and PI3K-mTOR mediate parallel pathways. , 1999, Molecular biology of the cell.

[4]  K. Vuori,et al.  The adaptor protein Crk connects multiple cellular stimuli to the JNK signaling pathway. , 1998, Proceedings of the National Academy of Sciences of the United States of America.

[5]  P. Vogt,et al.  The akt kinase: molecular determinants of oncogenicity. , 1998, Proceedings of the National Academy of Sciences of the United States of America.

[6]  S. Feller,et al.  Physiological signals and oncogenesis mediated through Crk family adapter proteins , 1998, Journal of cellular physiology.

[7]  J. Downward Mechanisms and consequences of activation of protein kinase B/Akt. , 1998, Current opinion in cell biology.

[8]  M. Karin,et al.  Regulation and function of the JNK subgroup of MAP kinases. , 1997, Biochimica et biophysica acta.

[9]  G. Panayotou,et al.  Phosphoinositide 3-kinases: a conserved family of signal transducers. , 1997, Trends in biochemical sciences.

[10]  H. Iba,et al.  Expression of a Constitutively Active Phosphatidylinositol 3-Kinase Induces Process Formation in Rat PC12 Cells , 1997, The Journal of Biological Chemistry.

[11]  L. Cantley,et al.  Transformation of chicken cells by the gene encoding the catalytic subunit of PI 3-kinase. , 1997, Science.

[12]  T. Ouchi,et al.  Downstream of Crk adaptor signaling pathway: activation of Jun kinase by v-Crk through the guanine nucleotide exchange protein C3G. , 1997, Proceedings of the National Academy of Sciences of the United States of America.

[13]  P. Warne,et al.  R-Ras can activate the phosphoinositide 3-kinase but not the MAP kinase arm of the Ras effector pathways , 1997, Current Biology.

[14]  H. Greulich,et al.  A role for Ras in v-Crk transformation. , 1996, Cell growth & differentiation : the molecular biology journal of the American Association for Cancer Research.

[15]  J. Guan,et al.  Phosphorylation of Tyrosine 397 in Focal Adhesion Kinase Is Required for Binding Phosphatidylinositol 3-Kinase* , 1996, The Journal of Biological Chemistry.

[16]  R. Birge,et al.  SH2 and SH3‐containing adaptor proteins: redundant or independent mediators of intracellular signal transduction , 1996, Genes to cells : devoted to molecular & cellular mechanisms.

[17]  K. Shimotohno,et al.  Expression of cell-cycle regulatory genes in HTLV-I infected T-cell lines: possible involvement of Tax1 in the altered expression of cyclin D2, p18Ink4 and p21Waf1/Cip1/Sdi1. , 1996, Oncogene.

[18]  B. Mayer,et al.  Differential inhibition of signaling pathways by dominant-negative SH2/SH3 adapter proteins , 1995, Molecular and cellular biology.

[19]  E. Goldsmith,et al.  How MAP Kinases Are Regulated (*) , 1995, The Journal of Biological Chemistry.

[20]  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.

[21]  Y. Yazaki,et al.  The C-terminal SH3 domain of the mouse c-Crk protein negatively regulates tyrosine-phosphorylation of Crk associated p130 in rat 3Y1 cells. , 1994, Oncogene.

[22]  M. Shibuya,et al.  C3G, a guanine nucleotide-releasing protein expressed ubiquitously, binds to the Src homology 3 domains of CRK and GRB2/ASH proteins. , 1994, Proceedings of the National Academy of Sciences of the United States of America.

[23]  K Y Hui,et al.  A specific inhibitor of phosphatidylinositol 3-kinase, 2-(4-morpholinyl)-8-phenyl-4H-1-benzopyran-4-one (LY294002). , 1994, The Journal of biological chemistry.

[24]  T. Okada,et al.  Essential role of phosphatidylinositol 3-kinase in insulin-induced glucose transport and antilipolysis in rat adipocytes. Studies with a selective inhibitor wortmannin. , 1994, The Journal of biological chemistry.

[25]  T. Pawson,et al.  SH2 and SH3 domains: From structure to function , 1992, Cell.

[26]  H. Hanafusa,et al.  Activation of c-Src in cells bearing v-Crk and its suppression by Csk , 1992, Molecular and cellular biology.

[27]  M. Shibuya,et al.  Two species of human CRK cDNA encode proteins with distinct biological activities , 1992, Molecular and cellular biology.

[28]  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.

[29]  H. Hanafusa,et al.  Biological and biochemical activity of v-Crk chimeras containing the SH2/SH3 regions of phosphatidylinositol-specific phospholipase C-gamma and Src , 1992, Journal of virology.

[30]  M. Boyd,et al.  Morphometric and colorimetric analyses of human tumor cell line growth and drug sensitivity in soft agar culture. , 1991, Cancer research.

[31]  B. Mayer,et al.  Mutagenic analysis of the v-crk oncogene: requirement for SH2 and SH3 domains and correlation between increased cellular phosphotyrosine and transformation , 1990, Journal of virology.

[32]  H. Hanafusa,et al.  Phosphatidylinositol kinase type I activity associates with various oncogene products. , 1989, Oncogene research.

[33]  B. Mayer,et al.  A novel viral oncogene with structural similarity to phospholipase C , 1988, Nature.

[34]  D. Boettiger,et al.  Epitope mapping of monoclonal antibodies to gag protein p19 of avian sarcoma and leukaemia viruses. , 1987, The Journal of general virology.

[35]  C. Stoltzfus,et al.  Gene expression from both intronless and intron-containing Rous sarcoma virus clones is specifically inhibited by anti-sense RNA , 1985, Molecular and cellular biology.

[36]  H. Hanafusa Rapid transformation of cells by Rous sarcoma virus. , 1969, Proceedings of the National Academy of Sciences of the United States of America.