Ret-mediated mitogenesis requires Src kinase activity.

The proto-oncogene RET encodes a transmembrane growth neurotrophic receptor with tyrosine kinase (TK) activity. RET mutations are associated with several human neoplastic and nonneoplastic diseases, including thyroid papillary carcinoma, multiple endocrine neoplasia type 2 syndromes, and Hirschsprung's disease. Activation of receptor TKs results in the binding and activation of downstream signaling proteins, among which are nonreceptor TKs of the Src family. To test the involvement of c-Src in Ret-mediated signaling, we measured the levels of c-Src activity in NIH3T3 cells coexpressing Ret and the accessory GFR alpha-1 receptor or an epidermal growth factor receptor/Ret chimeric receptor when the cells were stimulated by glial cell line-derived neurotrophic factor or epidermal growth factor, respectively. Ret stimulation resulted in the activation of c-Src. We also measured the levels of Src kinase activity in cell lines expressing isoforms of the Ret receptor activated by different mutations. These cells showed higher Src kinase activity than the normal counterpart. Furthermore, we show that Ret is able to associate with the SH2 domain of Src in a phosphotyrosine-dependent fashion. Microinjection of a kinase inactive mutant of c-Src blocked Ret-mediated mitogenic effect. These experiments demonstrate that activated Ret is able to bind and stimulate c-Src kinase and that Src activation is essential for the mitogenic activity of Ret.

[1]  G. D. Vita,et al.  Glial cell line-derived neurotrophic factor differentially stimulates ret mutants associated with the multiple endocrine neoplasia type 2 syndromes and Hirschsprung's disease. , 1998, Endocrinology.

[2]  M. Santoro,et al.  Signalling of the Ret receptor tyrosine kinase through the c-Jun NH2-terminal protein kinases (JNKs): evidence for a divergence of the ERKs and JNKs pathways induced by Ret , 1998, Oncogene.

[3]  Susan S. Taylor,et al.  Shc and Enigma Are Both Required for Mitogenic Signaling by Ret/ptc2 , 1998, Molecular and Cellular Biology.

[4]  T. Boone,et al.  GFRα-2 and GFRα-3 Are Two New Receptors for Ligands of the GDNF Family , 1997, The Journal of Biological Chemistry.

[5]  T. Pawson,et al.  Signaling through scaffold, anchoring, and adaptor proteins. , 1997, Science.

[6]  Hua Yu,et al.  Activation of c-Src by receptor tyrosine kinases in human colon cancer cells with high metastatic potential , 1997, Oncogene.

[7]  Jimi Adu,et al.  Neurturin responsiveness requires a GPI-linked receptor and the Ret receptor tyrosine kinase , 1997, Nature.

[8]  Barbara Moffat,et al.  A GPI-linked protein that interacts with Ret to form a candidate neurturin receptor , 1997, Nature.

[9]  T. Pawson,et al.  RET alternate splicing influences the interaction of activated RET with the SH2 and PTB domains of Shc, and the SH2 domain of Grb2 , 1997, Oncogene.

[10]  M. Borrello,et al.  Identification of Shc docking site on Ret tyrosine kinase , 1997, Oncogene.

[11]  G. Botti,et al.  Development of mammary and cutaneous gland tumors in transgenic mice carrying the RET/PTC1 oncogene. , 1996, Oncogene.

[12]  N. Asai,et al.  A Mutation at Tyrosine 1062 in MEN2A-Ret and MEN2B-Ret Impairs Their Transforming Activity and Association with Shc Adaptor Proteins* , 1996, The Journal of Biological Chemistry.

[13]  G. D. Vita,et al.  A potential pathogenetic mechanism for multiple endocrine neoplasia type 2 syndromes involves ret-induced impairment of terminal differentiation of neuroepithelial cells. , 1996, Proceedings of the National Academy of Sciences of the United States of America.

[14]  T. Hunter,et al.  Requirement for c-Src Catalytic Activity and the SH3 Domain in Platelet-derived Growth Factor BB and Epidermal Growth Factor Mitogenic Signaling* , 1996, The Journal of Biological Chemistry.

[15]  S. Courtneidge,et al.  The Src SH3 Domain Is Required for DNA Synthesis Induced by Platelet-derived Growth Factor and Epidermal Growth Factor* , 1996, The Journal of Biological Chemistry.

[16]  J. Louis,et al.  GDNF–Induced Activation of the Ret Protein Tyrosine Kinase Is Mediated by GDNFR-α, a Novel Receptor for GDNF , 1996, Cell.

[17]  Vittorio,et al.  Molecular heterogeneity of RET loss of function in Hirschsprung's disease. , 1996, The EMBO journal.

[18]  Susan S. Taylor,et al.  Mitogenic Signaling by Ret/ptc2 Requires Association with Enigma via a LIM Domain* , 1996, The Journal of Biological Chemistry.

[19]  Pier Paolo Di Fiore,et al.  Direct Association between the Ret Receptor Tyrosine Kinase and the Src Homology 2-containing Adapter Protein Grb7 (*) , 1996, The Journal of Biological Chemistry.

[20]  M. Borrello,et al.  The full oncogenic activity of Ret/ptc2 depends on tyrosine 539, a docking site for phospholipase Cgamma , 1996, Molecular and cellular biology.

[21]  B. Pasini,et al.  RET mutations in human disease. , 1996, Trends in genetics : TIG.

[22]  R. A. Decker,et al.  Oncogenic RET Receptors Display Different Autophosphorylation Sites and Substrate Binding Specificities (*) , 1996, The Journal of Biological Chemistry.

[23]  J. Bos,et al.  Ret receptor tyrosine kinase activates extracellular signal-regulated kinase 2 in SK-N-MC cells. , 1995, Oncogene.

[24]  M. Barone,et al.  Myc but not Fos rescue of PDGF signalling block caused by kinase-inactive Src , 1995, Nature.

[25]  M. Santoro,et al.  Molecular defects in thyroid carcinomas: role of the RET oncogene in thyroid neoplastic transformation. , 1995, European journal of endocrinology.

[26]  M. Santoro,et al.  Expression of the RET oncogene induces differentiation of SK-N-BE neuroblastoma cells. , 1995, Cell growth & differentiation : the molecular biology journal of the American Association for Cancer Research.

[27]  Susan S. Taylor,et al.  Tyrosines outside the Kinase Core and Dimerization Are Required for the Mitogenic Activity of RET/ptc2 (*) , 1995, The Journal of Biological Chemistry.

[28]  A. Pandey,et al.  The Ret Receptor Protein Tyrosine Kinase Associates with the SH2-containing Adapter Protein Grb10 (*) , 1995, The Journal of Biological Chemistry.

[29]  W. Muller,et al.  Direct and specific interaction of c-Src with Neu is involved in signaling by the epidermal growth factor receptor. , 1995, Oncogene.

[30]  G. D. Vita,et al.  Activated RET/PTC oncogene elicits immediate early and delayed response genes in PC12 cells. , 1995, Oncogene.

[31]  M. Borrello,et al.  Loss of function effect of RET mutations causing Hirschsprung disease , 1995, Nature Genetics.

[32]  M. Roussel,et al.  DNA synthesis induced by some but not all growth factors requires Src family protein tyrosine kinases , 1995, Molecular and cellular biology.

[33]  al. et,et al.  Activation of RET as a dominant transforming gene by germline mutations of MEN2A and MEN2B , 1995, Science.

[34]  A Greco,et al.  The oncogenic versions of the Ret and Trk tyrosine kinases bind Shc and Grb2 adaptor proteins. , 1994, Oncogene.

[35]  L. Philipson,et al.  CHOP (GADD153) and its oncogenic variant, TLS-CHOP, have opposing effects on the induction of G1/S arrest. , 1994, Genes & development.

[36]  B. Ponder,et al.  Mutations of the RET proto-oncogene in Hirschsprung's disease , 1994, Nature.

[37]  B. Pasini,et al.  Point mutations affecting the tyrosine kinase domain of the RET proto-oncogene in Hirschsprung's disease , 1994, Nature.

[38]  M. Luther,et al.  Involvement of pp60c-src with two major signaling pathways in human breast cancer. , 1994, Proceedings of the National Academy of Sciences of the United States of America.

[39]  M. Santoro,et al.  An epidermal growth factor receptor/ret chimera generates mitogenic and transforming signals: evidence for a ret-specific signaling pathway , 1994, Molecular and cellular biology.

[40]  W. Muller,et al.  Mammary tumors expressing the neu proto-oncogene possess elevated c-Src tyrosine kinase activity , 1994, Molecular and cellular biology.

[41]  Jonathan A. Cooper,et al.  The when and how of Src regulation , 1993, Cell.

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

[43]  M. Roussel,et al.  Activation of Src family kinases by colony stimulating factor‐1, and their association with its receptor. , 1993, The EMBO journal.

[44]  M. Santoro,et al.  The TRK and RET tyrosine kinase oncogenes cooperate with ras in the neoplastic transformation of a rat thyroid epithelial cell line. , 1993, Cell growth & differentiation : the molecular biology journal of the American Association for Cancer Research.

[45]  G. Staal,et al.  Characterization of protein tyrosine kinases from human breast cancer: involvement of the c-src oncogene product. , 1992, Cancer research.

[46]  J. Hoxie,et al.  Activation of Fc gamma RII induces tyrosine phosphorylation of multiple proteins including Fc gamma RII. , 1992, The Journal of biological chemistry.

[47]  S. Courtneidge,et al.  Association of Fyn with the activated platelet-derived growth factor receptor: requirements for binding and phosphorylation. , 1992, Oncogene.

[48]  J. Brugge,et al.  Signal transduction by nerve growth factor and fibroblast growth factor in PC12 cells requires a sequence of src and ras actions , 1991, The Journal of cell biology.

[49]  S. Courtneidge,et al.  Association between the PDGF receptor and members of the src family of tyrosine kinases , 1990, Cell.

[50]  S. Aaronson,et al.  EGF receptor and erbB-2 tyrosine kinase domains confer cell specificity for mitogenic signaling. , 1990, Science.

[51]  M. Santoro,et al.  PTC is a novel rearranged form of the ret proto-oncogene and is frequently detected in vivo in human thyroid papillary carcinomas , 1990, Cell.

[52]  J. Pipas,et al.  pp60c-src activation in human colon carcinoma. , 1989, The Journal of clinical investigation.

[53]  T. Iwamoto,et al.  Cloning and expression of the ret proto-oncogene encoding a tyrosine kinase with two potential transmembrane domains. , 1988, Oncogene.

[54]  B. Groner,et al.  Prolactin regulation of beta‐casein gene expression and of a cytosolic 120‐kd protein in a cloned mouse mammary epithelial cell line. , 1988, The EMBO journal.

[55]  A. Fusco,et al.  One- and two-step transformations of rat thyroid epithelial cells by retroviral oncogenes , 1987, Molecular and cellular biology.

[56]  C R King,et al.  erbB-2 is a potent oncogene when overexpressed in NIH/3T3 cells. , 1987, Science.

[57]  P. Cohen,et al.  Analysis of pp60c-src protein kinase activity in human tumor cell lines and tissues. , 1986, The Journal of biological chemistry.

[58]  H. Rübsamen,et al.  Expression of pp60c-src protein kinase in adult and fetal human tissue: high activities in some sarcomas and mammary carcinomas. , 1983, Cancer research.