Receptor Tyrosine Kinase (RTK) signaling is essential for normal biological processes and disruption of this regulation can lead to
tumor initiation and progression. Cbl
proteins (Cbl, Cbl-b and Cbl-c) are a family of RING finger (RF)
ubiquitin ligases that negatively regulate a variety of RTKs, including EGFR, MET, and RET. Recent studies have identified Cbl mutations associated with human myeloid
neoplasias in approximately 5% of the cases. Cbl-c is the most recently identified
human Cbl protein and is expressed exclusively in epithelial cells. We identified a novel
cDNA that was isolated from a mouse
mammary cancer from the C3(1)
Large T Antigen transgenic model. This mutant
cDNA encodes a
protein that has a deletion in the RF domain of Cbl-c, thereby resembling known Cbl family mutations associated with myeoloid
neoplasias. Genomic analysis of both parental and transgenic lines shows no evidence of germline mutation indicating that this mutation is likely a somatic mutation. The
mutant protein enhances transformation of NIH 3T3 cells when expressed in combination with SV40
Large T antigen. Together these data are consistent with a second hit mutation. In overexpression studies, this mutant Cbl-c
protein fails to mediate ubiquitination of activated EGFR and acts in a dominant negative fashion to prevent ubiquitination and downregulation of the activated EGFR by wild type Cbl
proteins. Mechanistically, the mutant Cbl-c binds to the EGFR and prevents recruitment of the wild type Cbl
protein. Furthermore, data mining reveals Cbl-c mutations associated with solid
tumors in humans. Subsequent cell-based analysis demonstrates a similar loss of E3 function and dominant negative effects for one of these human mutations. These data suggest that like Cbl mutations in myeloid
neoplasms, loss of Cbl-c function may contribute to the pathogenesis of solid
tumors in murine models and in humans.