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Cbl-b interacts with ubiquitinated proteins; differential functions of the UBA domains of c-Cbl and Cbl-b

Oncogene volume 23, pages 7104–7115 (2004)Cite this article

Abstract

Cbl proteins are ubiquitin protein ligases, which ubiquitinate activated tyrosine kinases and target them for degradation. Both c-Cbl and Cbl-b have an ubiquitin associated (UBA) domain at their C-terminal end. We observed that high molecular weight ubiquitinated proteins constitutively coimmunoprecipitated with transfected and endogenous Cbl-b, but not c-Cbl. The binding site for these ubiquitinated proteins was mapped to the UBA domain of Cbl-b (UBAb). GST-fusion proteins containing the UBAb interacted with ubiquitinated proteins and polyubiquitin chains in vitro, whereas those containing the UBA domain of c-Cbl (UBAc) did not. The UBAb had a much greater affinity for polyubiquitin chains than for monoubiquitin. Analysis of the UBAb and UBAc demonstrate that the affinity for ubiquitin is determined by multiple amino-acid differences between the two domains. Overexpression of the UBAb, but not overexpression of the UBAc, inhibited a variety of ubiquitin-mediated processes such as degradation of ubiquitinated proteins (i.e. EGFR, Mdm-2, and Siah-1). This in vivo result is consistent with the differences in ubiquitin binding observed in vitro between the UBAb and UBAc. This difference in ubiquitin-binding may reflect distinct regulatory functions of c-Cbl and Cbl-b.

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Acknowledgements

We thank Lawrence Samelson and Alan Weissman for helpful discussion and critical review of the manuscript.

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Author notes

  1. Gareth C Davies and Seth A Ettenberg: GCD and SAE contributed equally to this work.

Authors and Affiliations

  1. Laboratory of Cellular and Molecular Biology, Center for Cancer Research, Bethesda, 20892, MD, USA

    Gareth C Davies, Seth A Ettenberg, Ashley O Coats, Mark Mussante, Marion M Nau & Stan Lipkowitz

  2. Advanced Biomedical Computing Center, National Cancer Institute, National Institutes of Health, Bethesda, 20892, MD, USA

    Sarangan Ravichandran & Jack Collins

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  1. Gareth C Davies
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Correspondence to Stan Lipkowitz.

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Davies, G., Ettenberg, S., Coats, A. et al. Cbl-b interacts with ubiquitinated proteins; differential functions of the UBA domains of c-Cbl and Cbl-b. Oncogene 23, 7104–7115 (2004). https://doi.org/10.1038/sj.onc.1207952

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