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Tyrosine phosphorylation of C-Cbl facilitates adhesion and spreading while suppressing anchorage-independent growth of V-Abl-transformed NIH3T3 fibroblasts

Oncogene volume 18, pages 3703–3715 (1999)Cite this article

Abstract

The protooncogenic protein c-Cbl becomes tyrosine phosphorylated in normal cells in response to a variety of external stimuli, as well as in cells transformed by oncogenic protein tyrosine kinases. Tyrosine phosphorylation of c-Cbl upregulates its binding to multiple crucial signaling molecules. However, the biological consequences of c-Cbl-mediated signaling are insufficiently understood. To analyse the biological functions of c-Cbl, we overexpressed wild-type c-Cbl and its tyrosine phosphorylation-defective mutant form in v-Abl-transformed NIH3T3 fibroblasts. In this system, wild-type c-Cbl facilitated adhesion and spreading of v-Abl-transformed fibroblasts on the extracellular matrix, while reducing anchorage independence of these cells, as measured by their colony-forming efficiency in soft agar. Therefore, overexpression of wild-type c-Cbl exhibits an overall transformation-suppressing effect. By contrast, overexpression of a tyrosine phosphorylation-defective form of c-Cbl increases neither adhesion nor anchorage dependence of v-Abl-transformed fibroblasts. Analysis of the role of individual tyrosine phosphorylation sites of c-Cbl in these phenomena indicates that both phosphatidylinositol-3′ kinase and the CrkL adaptor protein may be involved in the observed effects of c-Cbl. To summarize, the results presented in this report indicate that c-Cbl is involved in regulation of cell adhesion and cytoskeletal rearrangements, and that these effects of c-Cbl are dependent on its tyrosine phosphorylation.

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Abbreviations

HA:

hemagglutinin

mAb:

monoclonal antibody

PBS:

phosphate-buffered saline

PI-3′ kinase:

phosphatidylinositol-3′ kinase

PTK:

protein tyrosine kinase

P-Tyr:

phosphotyrosine

SDS – PAGE:

sodium dodecyl sulfate-polyacrylamine gel electrophoresis

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Acknowledgements

We thank R Hawley, A Kingsman, and W Langdon for providing reagents, A Teckchandani for help with some experiments, M Blake for help with phase-contrast microscopy, J Gibas for help with confocal microscopy, and G Harvey for editorial help. This work was supported by an award from the Council for Tobacco Research USA and Public Health Service grant CA78499 (both to AY Tsygankov).

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  1. Department of Microbiology and Immunology, Temple University School of Medicine, 3400 Broad Street, Philadelphia, PA 19140, Pennsylvania, USA

    Elena A Feshchenko & Alexander Y Tsygankov

  2. The Fels Institute for Cancer Research and Molecular Biology, Temple University School of Medicine, 3400 Broad Street, Philadelphia, PA 19140, Pennsylvania, USA

    Scott K Shore

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Feshchenko, E., Shore, S. & Tsygankov, A. Tyrosine phosphorylation of C-Cbl facilitates adhesion and spreading while suppressing anchorage-independent growth of V-Abl-transformed NIH3T3 fibroblasts. Oncogene 18, 3703–3715 (1999). https://doi.org/10.1038/sj.onc.1202672

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