Isolation and characterization of a novel, transforming allele of the c-Cbl proto-oncogene from a murine macrophage cell line.
The c-Cbl proto-oncogene acts as an E3 ubiquitin ligase via its RING finger domain to negatively regulate activated cellular signal transduction pathways. We have identified an aberrant Cbl-protein of approximately 95 kDa, which we have called p95Cbl, from the murine reticulum sarcoma cell-line, J-774. Cloning of the p95Cbl cDNA revealed that it contains a deletion resulting in the loss of 111 amino acids, eliminating two critical tyrosine residues in the linker region as well as the entire RING finger domain. p95Cbl displays a propensity for its interaction with the Src-family kinase Hck over cellular Cbl expressed in the same cells. Like its wildtype counterpart, p95Cbl is inducibly tyrosine phosphorylated in response to Fcgamma receptor engagement on hematopoietic cells, however this phosphorylation is sustained beyond that of cellular Cbl. NIH3T3 fibroblasts stably expressing p95Cbl acquire the typical refractile morphology associated with cellular transformation and form colonies in a focus-formation assay. The exogenously expressed mutant protein is constitutively phosphorylated in fibroblasts and partitions into the particulate fraction of cells, while cellular Cbl is exclusively cytoplasmic. p95Cbl is a novel, oncogenic mutant of the c-Cbl proto-oncogene, which might act in a dominant negative fashion to prolong normal cellular signaling responses by interfering with the down-regulation of activated signaling complexes through c-Cbl.
Bisson
Sabine A
SA
Department of Oncology, The University of Calgary, 3330 Hospital Drive NW Calgary, Alberta T2N-4N1, Canada.
Ujack
Eva E
EE
Robbins
Stephen M
SM
D016475:3T3 Cells; D000595:Amino Acid Sequence; D000818:Animals; D015153:Blotting, Western; D002460:Cell Line; D002471:Cell Transformation, Neoplastic; D005982:Glutathione Transferase / Q000378:metabolism; D006801:Humans; D008264:Macrophages; D051379:Mice; D008969:Molecular Sequence Data; D010766:Phosphorylation; D011505:Protein-Tyrosine Kinases / Q000378:metabolism; D000090063:Proto-Oncogene Mas; D011518:Proto-Oncogene Proteins / Q000737:chemistry / Q000235:genetics* / Q000302:isolation & purification* / Q000378:metabolism; D050721:Proto-Oncogene Proteins c-cbl; D051572:Proto-Oncogene Proteins c-hck; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D017452:Receptors, IgG / Q000378:metabolism; D044767:Ubiquitin-Protein Ligases
C000715470:MAS1 protein, human; D000090063:Proto-Oncogene Mas; D011518:Proto-Oncogene Proteins; D012333:RNA, Messenger; D017452:Receptors, IgG; D050721:Proto-Oncogene Proteins c-cbl; D044767:Ubiquitin-Protein Ligases; D005982:Glutathione Transferase; D011505:Protein-Tyrosine Kinases; C496631:HCK protein, human; C496632:Hck protein, mouse; D051572:Proto-Oncogene Proteins c-hck; C067419:CBL protein, human; C493849:Cbl protein, mouse