Abstract
Notch3 and pTα signaling events are essential for T-cell leukemogenesis and characterize murine and human T-cell acute lymphoblastic leukemia. Genetic ablation of pTα expression in Notch3 transgenic mice abrogates tumor development, indicating that pTα signaling is crucial to the Notch3-mediated leukemogenesis. Here we report a novel direct interaction between Notch3 and pTα. This interaction leads to the recruitment and persistence of the E3 ligase protein c-Cbl to the lipid rafts in Notch3-IC transgenic thymocytes. Conversely, deletion of pTα in Notch3 transgenic mice leads to cytoplasmic retention of c-Cbl that targets Notch3 protein to the proteasomal-degradative pathway. It appears that protein kinase C θ (PKCθ), by regulating tyrosine and serine phosphorylation of Cbl, is able to control its function. We report here that the increased Notch3-IC degradation correlates with higher levels of c-Cbl tyrosine phosphorylation in Notch3-IC/pTα−/− double-mutant thymocytes, which also display a decreased PKCθ activity. Our data indicate that pTα/pre-T-cell receptor is able to regulate the different subcellular localization of c-Cbl and, by regulating PKCθ activity, is also able to influence its ubiquitin ligase activity upon Notch3 protein.
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Acknowledgements
We thank Dr A Joutel for Notch3 antibodies and Dr R Bei for purification of mouse monoclonal pTα antibody, Dr F Grassi for c-Cbl plasmid and Dr M Bouché for PKCθ plasmids. This work was supported by the Italian Association for Cancer Research (AIRC), the Italian Ministry of University and Research (MIUR), PRIN Program, the Italian Ministry of Health and the European Union (Euro-Thymaide FP6 Integrated Project; LHSB-CT-2003-543410).
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Supplementary Information accompanies the paper on the Oncogene website (http://www.nature.com/onc)
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Checquolo, S., Palermo, R., Cialfi, S. et al. Differential subcellular localization regulates c-Cbl E3 ligase activity upon Notch3 protein in T-cell leukemia. Oncogene 29, 1463–1474 (2010). https://doi.org/10.1038/onc.2009.446
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DOI: https://doi.org/10.1038/onc.2009.446
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