Membrane-anchored Cbl suppresses Hck protein-tyrosine kinase mediated cellular transformation.
The mammalian proto-oncogene Cbl and its cellular homologues in Caenorhabditis elegans (Sli-1) and Drosophila (D-Cbl) are negative regulators of some growth factor receptor signaling pathways. Herein we show that Cbl can negatively regulate another signaling molecule, namely theSrc-family kinase Hck by targeting it for degradation. Hck-mediated cellular transformation of murine fibroblasts is reverted by ectopic expression of a membrane-anchored allele of Cbl as assessed by the cellular morphology, suppression of anchorage independent growth, and an overall reduction in the total tyrosine phosphorylation levels within the cells. The expression of Cbl at the plasma membrane targets both Hck and itself for ubiquitination and degradation, requiring an intact RING finger. Pharmacological inhibition of the proteasome prevents the degradation of Hck correlating with an increase in the phosphotyrosine levels within the cells. Activated Hck and membrane-anchored Cbl are present in similar subcellular localizations and co-immunoprecipitate, suggesting that their interaction is required for subsequent ubiquitination and degradation. Interestingly, both constitutively active and kinase-inactive Hck interact with and are targeted for degradation by Cbl. This work illustrates alternate means to regulate Src-family kinases, and suggests that Cbl may be able to suppress many signaling pathways that are activated in various proliferative syndromes including cancer.
Howlett
Christopher J
CJ
Department of Oncology, The University of Calgary, 3330 Hospital Drive N.W., Calgary, Alberta T2N-4N1, Canada.
Robbins
Stephen M
SM
D016475:3T3 Cells; D000818:Animals; D002471:Cell Transformation, Neoplastic; D003546:Cysteine Endopeptidases / Q000502:physiology; D051379:Mice; D009097:Multienzyme Complexes / Q000502:physiology; D046988:Proteasome Endopeptidase Complex; D011505:Protein-Tyrosine Kinases / Q000032:analysis / Q000037:antagonists & inhibitors* / Q000378:metabolism; D011518:Proto-Oncogene Proteins / Q000032:analysis / Q000037:antagonists & inhibitors* / Q000378:metabolism / Q000502:physiology*; D050721:Proto-Oncogene Proteins c-cbl; D051572:Proto-Oncogene Proteins c-hck; D025801:Ubiquitin / Q000378:metabolism; D044767:Ubiquitin-Protein Ligases
D009097:Multienzyme Complexes; D011518:Proto-Oncogene Proteins; D025801:Ubiquitin; D050721:Proto-Oncogene Proteins c-cbl; D044767:Ubiquitin-Protein Ligases; D011505:Protein-Tyrosine Kinases; C496632:Hck protein, mouse; D051572:Proto-Oncogene Proteins c-hck; D003546:Cysteine Endopeptidases; D046988:Proteasome Endopeptidase Complex; C493849:Cbl protein, mouse