The SH2-containing inositol 5-phosphatase (SHIP)-1 is implicated in the control of cell-cell junction and induces dissociation and dispersion of MDCK cells.

Hepatocyte growth factor (HGF) induces the breakdown of cell junction and the dispersion of colonies of epithelial cells, providing a model system for the investigation of the molecular mechanisms of one of the important aspects of tumorogenesis. We have previously reported that the SH2-domain-containing inositol 5'phosphatase (SHIP)-1 binds to c-Met, and potentiated HGF-mediated branching tubulogenesis. In this study, we describe the establishment of MDCK cell lines which express MycHis-tagged SHIP-1 at different levels. Expression of SHIP-1 in MDCK cells at a high level resulted in cell morphology characteristic of an epithelial-mesenchymal like transition; cells lost cortical actin, developed actin stress fibers and gained spontaneous motility without treatment of HGF. When the level of MycHis-tagged SHIP-expression was relatively low, transfectants partially lost cortical actin and phalloidin stained puncta appeared at cell-cell junctions even in the absence of HGF. The treatment of MAP kinase inhibitor, PD98059, did not influence SHIP-1 mediated alteration of adherens-junction of MDCK cells, while, phosphatidylinositol 3 (PI 3)- kinase inhibitor, LY294002, drastically reduced SHIP-1 mediated phenotype. Furthermore, expression of a mutant SHIP-1 lacking catalytic activity in MDCK cells did not alter the cortical actin distribution and HGF-mediated MAP and Akt kinase-phosphorylation, but suppressed HGF induced cell dispersion, suggesting that phosphatase activity is important for cytoskeleton rearrangement and cell dispersion.

c-Cbl associates directly with the C-terminal tail of the receptor for the macrophage colony-stimulating factor, c-Fms, and down-modulates this receptor but not the viral oncogene v-Fms.

The receptor for the macrophage colony-stimulating factor (CSF-1, also termed M-CSF), the tyrosine kinase c-Fms, was originally determined to be the oncogene product of the McDonough strain of feline sarcoma virus, v-Fms. The structural difference between c-Fms and v-Fms amounts to only five point mutations in the extracellular domain, two mutations in the cytoplasmic domain, and the replacement of 50 amino acids by 14 unrelated amino acids at the C-terminal tail. Here, we have identified c-Cbl as the direct binding partner for c-Fms. c-Cbl binds to phosphotyrosine residue 977 at the C-terminal end of feline c-Fms, which is absent in v-Fms. The replacement of the C-terminal end of v-Fms by the corresponding part of c-Fms (vc-Fms) restored the binding potential. As a result, vc-Fms reduced the transforming potency of v-Fms. The overexpression of Cbl did not influence the v-Fms-transformed phenotype, although c-Cbl forms a complex with v-Fms indirectly. In contrast, the expression of Cbl drastically reduced the vc-Fms-transformed phenotype and the activation of Erk and enhanced Fms ubiquitination via phosphotyrosine residue 977. Furthermore, the replacement of tyrosine 977 into phenylalanine in feline c-Fms and vc-Fms reduced the Cbl-dependent ubiquitination. These data suggest that an indirect association of c-Cbl via multimeric complex induced a different signaling pathway from the pathway induced by c-Cbl direct interaction.