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Structural basis for autoinhibition and phosphorylation-dependent activation of c-Cbl

Nature Structural & Molecular Biology volume 19, pages 184192 (2012) | Download Citation

Abstract

Cbls are RING ubiquitin ligases that attenuate receptor tyrosine kinase (RTK) signal transduction. Cbl ubiquitination activity is stimulated by phosphorylation of a linker helix region (LHR) tyrosine residue. To elucidate the mechanism of activation, we determined the structures of human CBL, a CBL−substrate peptide complex and a phosphorylated-Tyr371-CBL−E2−substrate peptide complex, and we compared them with the known structure of a CBL−E2−substrate peptide complex. Structural and biochemical analyses show that CBL adopts an autoinhibited RING conformation, where the RING's E2-binding surface associates with CBL to reduce E2 affinity. Tyr371 phosphorylation activates CBL by inducing LHR conformational changes that eliminate autoinhibition, flip the RING domain and E2 into proximity of the substrate-binding site and transform the RING domain into an enhanced E2-binding module. This activation is required for RTK ubiquitination. Our results present a mechanism for regulation of c-Cbl's activity by autoinhibition and phosphorylation-induced activation.

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Acknowledgements

We would like to thank B. Schulman, F. Kozielski and A. Schuettelkopf for helpful discussions; V. Ulaganathan for computer support; S. Lilla and N. Morrice for mass spectroscopic analyses; W. Clark and A. Keith for in-house DNA sequencing; the Diamond Light Source (DLS) for access to beamlines I02, I03 and I04−1 (mx1229 and mx6683) that contributed to the results presented here; and the ID14−1 beamline at the European Synchrotron Radiation Facility (ESRF) for access and synchrotron support. This work was supported by Cancer Research UK.

Author information

Author notes

    • Hao Dou
    •  & Lori Buetow

    These authors contributed equally to this work.

Affiliations

  1. The Beatson Institute for Cancer Research, Glasgow, UK.

    • Hao Dou
    • , Lori Buetow
    • , Andreas Hock
    • , Gary J Sibbet
    • , Karen H Vousden
    •  & Danny T Huang

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Contributions

H.D., L.B., G.J.S. and D.T.H. designed, conducted and analyzed in vitro experiments. A.H. designed, conducted and analyzed in vivo experiments. K.H.V. designed and analyzed in vivo experiments. L.B. and D.T.H. wrote the manuscript. All authors discussed the results and commented on the manuscript.

Competing interests

The authors declare no competing financial interests.

Corresponding author

Correspondence to Danny T Huang.

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DOI

https://doi.org/10.1038/nsmb.2231

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