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Crystal structure of the prefusion surface glycoprotein of the prototypic arenavirus LCMV

Nature Structural & Molecular Biology volume 23, pages 513521 (2016) | Download Citation

Abstract

Arenaviruses exist worldwide and can cause hemorrhagic fever and neurologic disease. A single glycoprotein expressed on the viral surface mediates entry into target cells. This glycoprotein, termed GPC, contains a membrane-associated signal peptide, a receptor-binding subunit termed GP1 and a fusion-mediating subunit termed GP2. Although GPC is a critical target of antibodies and vaccines, the structure of the metastable GP1–GP2 prefusion complex has remained elusive for all arenaviruses. Here we describe the crystal structure of the fully glycosylated prefusion GP1–GP2 complex of the prototypic arenavirus LCMV at 3.5 Å. This structure reveals the conformational changes that the arenavirus glycoprotein must undergo to cause fusion and illustrates the fusion regions and potential oligomeric states.

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Acknowledgements

The authors wish to acknowledge the Viral Hemorrhagic Fever Research Consortium and US National Institutes of Health grant 1U19AI109762-01 (E.O.S., J.E.R. and R.F.G.), US National Institutes of Health grant R21 AI116112 (E.O.S.), an Investigators in Pathogenesis of Infectious Diseases award from the Burroughs Wellcome Fund (E.O.S.) and US National Institutes of Health grants AI009484 (M.B.O.) and A1099699 (M.B.O.) for funding; X. Dai for assistance with data processing; M. Buchmeier (University of California, Irvine) for the anti-LCMV GP1 (KL25) antibody; S. Whelan (Harvard Medical School) for LAMP1-knockout cells; and C. Corbaci for assistance in creating figures. The authors also acknowledge beamlines 11-1, 12-1 and 12-2 of the Stanford Synchrotron Radiation Lightsource; 5.0.2, 5.0.3 and 8.2.2 of the Advanced Light Source; 19-ID, 23-ID-B and 23-ID-D of the Advanced Photon Source; and SOLEIL Proxima 1 (Gif-sur-Yvette, France) for data collection. This is manuscript #29106 from The Scripps Research Institute.

Author information

Author notes

    • Sébastien Igonet

    Present address: Calixar, Lyon, France.

    • Kathryn M Hastie
    •  & Sébastien Igonet

    These authors contributed equally to this work.

Affiliations

  1. Department of Immunology and Microbial Science, The Scripps Research Institute, La Jolla, California, USA.

    • Kathryn M Hastie
    • , Sébastien Igonet
    • , Brian M Sullivan
    • , Michelle A Zandonatti
    • , Michael B Oldstone
    •  & Erica Ollmann Saphire
  2. SOLEIL Synchrotron, Gif-sur-Yvette, France.

    • Pierre Legrand
  3. Department of Pediatrics, School of Medicine, Tulane University, New Orleans, Louisiana, USA.

    • James E Robinson
  4. Department of Microbiology and Immunology, Tulane University, New Orleans, Louisiana, USA.

    • Robert F Garry
  5. De´partement de Virologie, Unite´ de Virologie Structurale, Institut Pasteur, Paris, France.

    • Félix A Rey
  6. Skaggs Institute for Chemical Biology, The Scripps Research Institute, La Jolla, California, USA.

    • Erica Ollmann Saphire

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Contributions

K.M.H. built and refined the model, cloned the constructs for biochemical characterization, performed the receptor binding experiments, analyzed the data and wrote the manuscript; S.I. cloned the constructs for crystallization, produced and crystallized the recombinant GP ectodomains, collected diffraction data at room temperature, phased the data and determined the structure; B.M.S. grew the recombinant viruses, designed and performed the receptor binding experiments and analyzed the data; P.L. collected diffraction data on frozen crystals and built and refined the model; M.A.Z. cloned the constructs and produced recombinant protein; J.E.R. and R.F.G. generated and produced antibodies used throughout the studies; F.A.R. supervised the initial work toward structure determination; M.B.O. designed experiments and analyzed the data; and E.O.S. analyzed the data and wrote the manuscript. K.M.H. and S.I. contributed equally to the study. All authors commented on the manuscript.

Competing interests

The authors declare no competing financial interests.

Corresponding author

Correspondence to Erica Ollmann Saphire.

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  1. 1.

    Supplementary Text and Figures

    Supplementary Figures 1–5 and Supplementary Table 1

  2. 2.

    Supplementary Data Set 1

    Subunit requirements for immunoprecipitation of α-dystroglycan

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DOI

https://doi.org/10.1038/nsmb.3210

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