Letter | Published:

Structure of the E6/E6AP/p53 complex required for HPV-mediated degradation of p53

Nature volume 529, pages 541545 (28 January 2016) | Download Citation

Abstract

The p53 pro-apoptotic tumour suppressor is mutated or functionally altered in most cancers. In epithelial tumours induced by ‘high-risk’ mucosal human papilloma viruses, including human cervical carcinoma and a growing number of head-and-neck cancers1, p53 is degraded by the viral oncoprotein E6 (ref. 2). In this process, E6 binds to a short leucine (L)-rich LxxLL consensus sequence within the cellular ubiquitin ligase E6AP3. Subsequently, the E6/E6AP heterodimer recruits and degrades p53 (ref. 4). Neither E6 nor E6AP are separately able to recruit p53 (refs 3, 5), and the precise mode of assembly of E6, E6AP and p53 is unknown. Here we solve the crystal structure of a ternary complex comprising full-length human papilloma virus type 16 (HPV-16) E6, the LxxLL motif of E6AP and the core domain of p53. The LxxLL motif of E6AP renders the conformation of E6 competent for interaction with p53 by structuring a p53-binding cleft on E6. Mutagenesis of critical positions at the E6–p53 interface disrupts p53 degradation. The E6-binding site of p53 is distal from previously described DNA- and protein-binding surfaces of the core domain. This suggests that, in principle, E6 may avoid competition with cellular factors by targeting both free and bound p53 molecules. The E6/E6AP/p53 complex represents a prototype of viral hijacking of both the ubiquitin-mediated protein degradation pathway and the p53 tumour suppressor pathway. The present structure provides a framework for the design of inhibitory therapeutic strategies against oncogenesis mediated by human papilloma virus.

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Accessions

Primary accessions

Protein Data Bank

Data deposits

Coordinates and structure factors have been deposited in the Protein Data Bank (PDB) under accession number 4XR8.

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Acknowledgements

This work received institutional support from le Centre National de la Recherche Scientifique (CNRS), Université de Strasbourg, l’Institut National de la Santé et de la Recherche Médicale (INSERM) and Région Alsace. The work was supported by grants from Ligue contre le Cancer, National Institutes of Health (grant R01CA134737 to S.V.P.), l’Agence Nationale de la Recherche (ANR-13-JSV8-004-01), Instruct (ESFRI), the French Infrastructure for Integrated Structural Biology (FRISBI) and Fondation pour La Recherche Medicale (fellowship to F.X.R.F.). We thank P. Poussin-Courmontagne, E. Ennifar, V. Olieric and B. Kieffer for advice. The authors declare that the content is solely their responsibility and does not represent the official views of the National Institutes of Health.

Author information

Affiliations

  1. Equipe labellisée Ligue, Biotechnologie et signalisation cellulaire UMR 7242, Ecole Superieure de Biotechnologie de Strasbourg, Boulevard Sébastien Brant, BP 10413, F-67412 Illkirch, France

    • Denise Martinez-Zapien
    • , Juline Poirson
    • , Juan Ramirez
    • , Anne Forster
    • , Murielle Masson
    • , Gilles Travé
    •  & Katia Zanier
  2. Institut de Génétique et de Biologie Moléculaire et Cellulaire (IGBMC)/INSERM U964/CNRS UMR 7104/Université de Strasbourg, 1 rue Laurent Fries, BP 10142, F-67404 Illkirch, France

    • Francesc Xavier Ruiz
    • , André Mitschler
    • , Alexandra Cousido-Siah
    •  & Alberto Podjarny
  3. Department of Pathology, University of Virginia, PO Box 800904, Charlottesville, Virginia 22908-0904, USA

    • Scott Vande Pol

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Contributions

D.M.Z., J.P., A.M., J.R.R., A.F., A.C.S. and K.Z. performed experiments; F.X.R., A.P., D.M.Z. and K.Z. performed structure determination; D.M.Z., F.X.R., J.P., A.M., A.P. and K.Z. analysed the data; D.M.Z., J.P. and K.Z. prepared figures; K.Z. and GT wrote the manuscript together with comments from all authors; M.M., S.V.P., A.P., GT and K.Z. supervised the work.

Competing interests

The authors declare no competing financial interests.

Corresponding authors

Correspondence to Gilles Travé or Katia Zanier.

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    Supplementary Information

    This file contains Supplementary Methods.

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    Supplementary Figure 1

    This file contains the source gel data for Figure 3 and Extended Data Figure 8.

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https://doi.org/10.1038/nature16481

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