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Src-like adaptor protein regulates TCR expression on thymocytes by linking the ubiquitin ligase c-Cbl to the TCR complex

Nature Immunology volume 7, pages 57–66 (2006)Cite this article

Abstract

The adaptor molecule SLAP and E3 ubiquitin ligase c-Cbl each regulate expression of T cell receptor (TCR)–CD3 on thymocytes. Here we provide genetic and biochemical evidence that both molecules function in the same pathway. TCR-CD3 expression was similar in the absence of SLAP and/or c-Cbl. SLAP and c-Cbl were found to interact, and their expression together downregulated CD3ε. This required multiple domains in SLAP and the ring finger of c-Cbl. Furthermore, expression of SLAP and c-Cbl together induced TCRζ ubiquitination and degradation, preventing the accumulation of fully assembled recycling TCR complexes. These studies indicate that SLAP links the E3 ligase activity of c-Cbl to the TCR, allowing for stage-specific regulation of TCR expression.

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Figure 1: Sla−/−, Cbl−/− and Sla−/−Cbl−/− DP thymocytes have a similar phenotype.
Figure 2: Increased TCRζ expression and CD3ε recycling pool due to a defect in TCRζ degradation.
Figure 3: Coexpression of SLAP and c-Cbl downregulates CD3ε expression on Jurkat T cells.
Figure 4: CD3ε downregulation requires Lck.
Figure 5: TCR-CD3 downregulation requires multiple domains in SLAP and the ring finger of c-Cbl.
Figure 6: SLAP and c-Cbl target TCRζ for ubiquitination and degradation.

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Acknowledgements

We thank F. Brodsky, M. von Zastrow and members of the Weiss lab for comments and suggestions. Supported by the National Institutes of Health (CA72531 to A.W. and CA987986 to H.B.).

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Authors and Affiliations

  1. Department of Medicine, Rosalind Russell Medical Research Center for Arthritis, Howard Hughes Medical Institute, University of California San Francisco, San Francisco, 94143, California, USA

    Margaret D Myers & Arthur Weiss

  2. Howard Hughes Medical Institute and National Jewish Medical and Research Center, Denver, 80206, Colorado, USA

    Tomasz Sosinowski

  3. Division of Pediatric Immunology/Rheumatology, Department of Pediatrics, University of California San Francisco, San Francisco, 94143, California, USA

    Leonard L Dragone & Carmen White

  4. Division of Molecular Oncology, Department of Medicine, Evanston Northwestern Healthcare Research Institute, Feinberg School of Medicine,

    Hamid Band

  5. Department of Biochemistry, Molecular Biology & Cell Biology and Robert H. Lurie Comprehensive Cancer Center, Northwestern University, Evanston, 60201, Illinois, USA

    Hamid Band

  6. Microbiology Department, Columbia University, College of Physicians and Surgeons, New York, 10032, New York, USA

    Hua Gu

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  1. Margaret D Myers
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Correspondence to Arthur Weiss.

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Supplementary information

Supplementary Fig. 1

CD3ε internalization by DP thymocytes is normal in the absence of SLAP and/or c-Cbl. (PDF 184 kb)

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Myers, M., Sosinowski, T., Dragone, L. et al. Src-like adaptor protein regulates TCR expression on thymocytes by linking the ubiquitin ligase c-Cbl to the TCR complex. Nat Immunol 7, 57–66 (2006). https://doi.org/10.1038/ni1291

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