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Cryo-electron microscopy reconstruction of a poliovirus-receptor-membrane complex

Nature Structural & Molecular Biology volume 12, pages 615–618 (2005)Cite this article

Abstract

To study non-enveloped virus cell entry, a versatile in vitro model system was developed in which liposomes containing nickel-chelating lipids were decorated with His-tagged poliovirus receptors and bound to virus. This system provides an exciting opportunity for structural characterization of the early steps in cell entry in the context of a membrane. Here we report the three-dimensional structure of a poliovirus–receptor–membrane complex solved by cryo-electron microscopy (cryo-EM) at a resolution of 32 Å. Methods were developed to establish the symmetry of the complex objectively. This reconstruction demonstrates that receptor binding brings a viral five-fold axis close to the membrane. Density is clearly defined for the icosahedral virus, for receptors (including known glycosylation sites) and for the membrane bilayer. Apparent perturbations of the bilayer close to the viral five-fold axis may function in subsequent steps of cell entry.

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Figure 1: Poliovirus bound to receptor-decorated liposomes as visualized by cryo-EM.
Figure 2: Artificially dotted icosahedral reconstruction of a virus-receptor-liposome complex demonstrates that virus approaches membrane along its five-fold axis.
Figure 3: Five-fold symmetric three-dimensional reconstruction of native poliovirus bound to receptor-decorated liposomes.

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Acknowledgements

We thank S. Lanzavecchia for discussions, D. Gohara for rendering Figure 3d, T. Walz for access to instruments in the Harvard Medical School Electron Microscopy laboratory and Y. Cheng for instruction and assistance in their use. This work is supported by National Institutes of Health grant AI20566 (to J.M.H.) and National Science Foundation predoctoral fellowship (to D.B.).

Author information

Authors and Affiliations

  1. Committee on Higher Degrees in Biophysics, Harvard University, Cambridge, 02138, Massachusetts, USA

    Doryen Bubeck & James M Hogle

  2. Department of Biological Chemistry and Molecular Pharmacology, Harvard Medical School, Boston, 02115, Massachusetts, USA

    David J Filman & James M Hogle

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  1. Doryen Bubeck
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  2. David J Filman
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  3. James M Hogle
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Correspondence to James M Hogle.

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Supplementary information

Supplementary Fig. 1

Fourier shell correlation (FSC) for the final reconstruction of the poliovirus-receptor-liposome complex. (PDF 199 kb)

Supplementary Video 1

Fivefold-symmetric reconstruction of a poliovirus-receptor-liposome complex. The virus and receptor surfaces are green, and the membrane surface is purpose. The separation between these two regions was decided based on the 23 Å resolution cryo-EM structure of native poliovirus bound to sixty copies of soluble receptor1. In the movie, the membrane is displayed at a lower contour than the virus-receptor component. Note that at the lower contour level, continuous density is seen between receptors and the membrane surface virus-receptor component. (MOV 8969 kb)

Scene I: Side view of the virus-receptor-membrane complex rotating about its fivefold symmetry axis (vertical).

Scene II: Virus and receptor are viewed from the membrane-proximal side of the complex along the fivefold axis. Prominent glycosylation sites on the domain 2 of the receptor are yellow.

Scene III: Unobstructed view of the outer leaflet of the membrane. Near the fivefold axis, the membrane projects towards the virus. This apparent distortion may be involved in the formation of channels that are related to the translocation of viral RNA across the membrane2,3. The small spherical features, which appear at low contour, are located close to the predicted C termini of the soluble receptor1.

Scene IV: Truncation of the density with clipping planes allows the interior of the virion to be seen.

This animation was created by SciAna FilmWorks, http://www.scianafilms.com/

REFERENCES

1. Belnap, D.M. et al. Three-dimensional structure of poliovirus receptor bound to poliovirus. Proc. Natl. Acad. Sci. USA 97, 73-78 (2000). 2. Tosteson, M., Wang, H., Naumov, A. & Chow, M. Poliovirus binding to its receptor in lipid bilayers results in particle-specific, temperature-sensitive channels. J. Gen. Virol. 86, 1581-1589 (2004). 3. Danthi, P., Tosteson, M., Li., Q.H. & Chow, M. Genome delivery and ion channel properties are altered in VP4 mutants of poliovirus. J. Virol. 77, 5266-5274 (2003).

Supplementary Methods (PDF 36 kb)

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Bubeck, D., Filman, D. & Hogle, J. Cryo-electron microscopy reconstruction of a poliovirus-receptor-membrane complex. Nat Struct Mol Biol 12, 615–618 (2005). https://doi.org/10.1038/nsmb955

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