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Structural rearrangements in the membrane penetration protein of a non-enveloped virus

Nature volume 430, pages 1053–1058 (2004)Cite this article

Abstract

Non-enveloped virus particles (those that lack a lipid-bilayer membrane) must breach the membrane of a target host cell to gain access to its cytoplasm. So far, the molecular mechanism of this membrane penetration step has resisted structural analysis. The spike protein VP4 is a principal component in the entry apparatus of rotavirus, a non-enveloped virus that causes gastroenteritis and kills 440,000 children each year1. Trypsin cleavage of VP4 primes the virus for entry by triggering a rearrangement that rigidifies the VP4 spikes2. We have determined the crystal structure, at 3.2 Å resolution, of the main part of VP4 that projects from the virion. The crystal structure reveals a coiled-coil stabilized trimer. Comparison of this structure with the two-fold clustered VP4 spikes in a ∼12 Å resolution image reconstruction from electron cryomicroscopy of trypsin-primed virions shows that VP4 also undergoes a second rearrangement, in which the oligomer reorganizes and each subunit folds back on itself, translocating a potential membrane-interaction peptide from one end of the spike to the other. This rearrangement resembles the conformational transitions of membrane fusion proteins of enveloped viruses3,4,5,6.

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Figure 1: Rotavirus virion structure, VP4 domains and fragments.
Figure 2: VP5CT structure.
Figure 3: The VP5* antigen domain and the VP8* core fit to an electron cryomicroscopy image reconstruction.
Figure 4: Model for VP4 rearrangements during priming and entry.

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Acknowledgements

We thank M. Babyonyshev for technical assistance; T. Yeates for help in analysing the crystal twinning disorder; H. Greenberg for cloned genes and recombinant baculoviruses; E. Vogan for help with data collection and analysis; and the staff of Advanced Photon Source beamline ID-19 (Argonne National Laboratory) and Cornell High Energy Synchrotron Source beamlines F1 and A1. We acknowledge the use of electron cryomicroscopy facilities at the National Center for Macromolecular Imaging funded by NIH at Baylor College of Medicine. This work was supported by an NIH grant and an Ellison Medical Foundation New Investigator in Global Infectious Diseases award to P.R.D., by an NIH grant to B.V.V.P., and by an NIH grant to S.C.H., who is a Howard Hughes Medical Institute Investigator.

Author information

Authors and Affiliations

  1. Department of Pediatrics, Harvard Medical School, and the Laboratory of Molecular Medicine, Children's Hospital,

    Philip R. Dormitzer & Stephen C. Harrison

  2. Howard Hughes Medical Institute, 320 Longwood Avenue, Boston, Massachusetts, 02115, USA

    Stephen C. Harrison

  3. Verna and Marrs McLean Department of Biochemistry and Molecular Biology, Baylor College of Medicine, One Baylor Plaza, Houston, Texas, 77030, USA

    Emma B. Nason & B. V. Venkataram Prasad

Authors
  1. Philip R. Dormitzer
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  2. Emma B. Nason
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  4. Stephen C. Harrison
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Correspondence to Philip R. Dormitzer.

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The authors declare that they have no competing financial interests.

Supplementary information

Supplementary Figure 1

Comparison of the rotavirus VP4 F'G loop and the alphavirus E1 fusion loop. Includes an amino acid sequence comparison and a structural comparison. (PDF 158 kb)

Supplementary Figure 2

Sample of electron density. An image from an unaveraged simulated annealing omit map is shown. (PDF 1474 kb)

Supplementary Methods

Details of the crystallographic structure determination. Includes a description of the twinning disorder in the crystals. (DOC 35 kb)

Supplementary Table 1

Neutralization escape mutations. Mutations that map to the VP5* fragment are assigned to epitopes based on the VP5CT structure. (DOC 62 kb)

Supplementary Table 2

Rotavirus VP4 sequences used to assess variability and obtain a consensus sequence. The variability in these sequences is mapped onto the surfaces of the VP8* core and VP5* antigen domain in Fig. 3a, c. The consensus sequence is used to assess the similarity between the VP4 F'G loop and the alphavirus E1 fusion loop in Supplementary Fig. 1. (DOC 37 kb)

Supplementary Table 3

Alphavirus E1 sequences used to obtain a consensus. The consensus sequence is used to assess the similarity between the VP4 F'G loop and the alphavirus E1 fusion loop in Supplementary Fig. 1. (DOC 26 kb)

Supplementary Table 4

X-ray diffraction data collection, phasing, and refinement statistics. (DOC 47 kb)

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Dormitzer, P., Nason, E., Venkataram Prasad, B. et al. Structural rearrangements in the membrane penetration protein of a non-enveloped virus. Nature 430, 1053–1058 (2004). https://doi.org/10.1038/nature02836

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