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Glycoprotein organization of Chikungunya virus particles revealed by X-ray crystallography

Nature volume 468, pages 709–712 (2010)Cite this article

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Abstract

Chikungunya virus (CHIKV) is an emerging mosquito-borne alphavirus that has caused widespread outbreaks of debilitating human disease in the past five years1. CHIKV invasion of susceptible cells is mediated by two viral glycoproteins, E1 and E2, which carry the main antigenic determinants and form an icosahedral shell at the virion surface. Glycoprotein E2, derived from furin cleavage of the p62 precursor into E3 and E2, is responsible for receptor binding, and E1 for membrane fusion. In the context of a concerted multidisciplinary effort to understand the biology of CHIKV2, here we report the crystal structures of the precursor p62–E1 heterodimer and of the mature E3–E2–E1 glycoprotein complexes. The resulting atomic models allow the synthesis of a wealth of genetic, biochemical, immunological and electron microscopy data accumulated over the years on alphaviruses in general. This combination yields a detailed picture of the functional architecture of the 25 MDa alphavirus surface glycoprotein shell. Together with the accompanying report on the structure of the Sindbis virus E2–E1 heterodimer at acidic pH (ref. 3), this work also provides new insight into the acid-triggered conformational change on the virus particle and its inbuilt inhibition mechanism in the immature complex.

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Figure 1: Structure of the p62–E1 heterodimer.
Figure 2: The E1 fusion loop binding groove in E2.
Figure 3: Combination with cryo-EM data.
Figure 4: p62 maturation and low pH conformational transition.

Accession codes

Primary accessions

Protein Data Bank

Data deposits

Structure factors and coordinates for the structures of the p62–E1 (crystal IO), E3–E2–E1sp (crystal MM), E3–E2–E1f (crystal MO1), E3–E2–E1t (crystal MO2) and Os2-E3–E2–E1t (crystal MO3) complexes have been deposited in the Protein Data Bank under accession codes 3N40, 3N41, 3N42, 3N43 and 3N44, respectively. The coordinates of the molecules fitted in the cryo-EM 3D reconstructions of SINV and SFV particles were deposited under accession codes 2XFB and 2XFC, respectively.

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Acknowledgements

We thank the CHIKV task force at Institut Pasteur, in particular the group of F. Tangy and the staff of platform PF8 for the CHIKV complementary DNA; A. Haouz of PF6 for crystallogenesis; the staff of synchrotron beamlines PROXIMA 1 at Soleil, ID23-eh2 at the European Synchrotron Radiation Facility and PX-I at the Swiss Light Source; M. Rossmann and Y. Sun for providing the 16 Å cryo-EM map of CHIKV virion-like particles and for sharing the coordinates and manuscript of the low pH structure of the SINV E1–E2 heterodimer before publication; and members of the F.A.R. laboratory for help during data collection. J.E.V. was supported by a Marie Curie fellowship through the European Union Research Traning Network program “Intrapath”. This work was funded in part by the French ‘Agence Nationale de la Recherche’ grant DENtry in the program ‘Microbiologie, Infections et Immunité’, by Merck-Serono, by the Pediatric Dengue Vaccine Initiative and by the Institut Pasteur program PTR201 CHIKV to F.A.R..

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Authors and Affiliations

  1. Département de Virologie, Institut Pasteur, Unité de Virologie Structurale, 25 rue du Dr Roux, 75724 Paris Cedex 15, France,

    James E. Voss, Marie-Christine Vaney, Stéphane Duquerroy & Félix A. Rey

  2. CNRS URA 3015, 25 rue du Dr Roux, 75724 Paris Cedex 15, France ,

    James E. Voss, Marie-Christine Vaney, Stéphane Duquerroy & Félix A. Rey

  3. Université Paris-Sud, Faculté d’Orsay, Orsay Cedex, 91405, France

    Stéphane Duquerroy

  4. Global Phasing Ltd, Sheraton House, Castle Park, Cambridge, CB3 0AX, United Kingdom

    Clemens Vonrhein & Gérard Bricogne

  5. Département de Biologie Structurale et Chimie, Institut Pasteur, Plateforme de Production de protéines recombinantes, 25 rue du Dr Roux, 75724 Paris Cedex 15, France,

    Christine Girard-Blanc & Elodie Crublet

  6. CNRS URA 2185, 25 rue du Dr Roux, 75724 Paris Cedex 15, France ,

    Christine Girard-Blanc & Elodie Crublet

  7. Synchrotron SOLEIL, L’Orme de Merisiers, BP 48 St Aubin, 91192 Gif sur Yvette, France ,

    Andrew Thompson

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  1. James E. Voss
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Contributions

J.E.V. made the constructs, produced and purified the protein, grew the crystals and participated in diffraction data collection; analysed the literature and prepared tables and figures. M.-C.V. carried out most of the various crystallographic refinements and prepared the figures, S.D. carried out the fitting into the cryo-EM maps of various alphavirus particles and prepared the figures, C.G.-B. and E.C. participated in optimizing protein production in large scale for crystal trials, C.V. and G.B. participated in data processing and in the structure determination. A.T. carried out specific data collection strategies to improve the signal to noise to extract anomalous signal for phasing; F.A.R. conceived the experiments and wrote the manuscript.

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Correspondence to Félix A. Rey.

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Supplementary information

Supplementary Information

The file contains Supplementary Analysis, Supplementary References, Supplementary Tables 1-7 and Supplementary Figures 1-7 with legends. (ZIP 12830 kb)

Supplementary Movie 1

This movie shows Six spikes around a Q6 axis of the T=4 icosahedral lattice (see legend in Supplementary Information). (MOV 4582 kb)

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Voss, J., Vaney, MC., Duquerroy, S. et al. Glycoprotein organization of Chikungunya virus particles revealed by X-ray crystallography. Nature 468, 709–712 (2010). https://doi.org/10.1038/nature09555

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