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GM1 structure determines SV40-induced membrane invagination and infection

Nature Cell Biology volume 12, pages 11–18 (2010)Cite this article

Abstract

Incoming simian virus 40 (SV40) particles enter tight-fitting plasma membrane invaginations after binding to the carbohydrate moiety of GM1 gangliosides in the host cell plasma membrane through pentameric VP1 capsid proteins. This is followed by activation of cellular signalling pathways, endocytic internalization and transport of the virus via the endoplasmic reticulum to the nucleus. Here we show that the association of SV40 (as well as isolated pentameric VP1) with GM1 is itself sufficient to induce dramatic membrane curvature that leads to the formation of deep invaginations and tubules not only in the plasma membrane of cells, but also in giant unilamellar vesicles (GUVs). Unlike native GM1 molecules with long acyl chains, GM1 molecular species with short hydrocarbon chains failed to support such invagination, and endocytosis and infection did not occur. To conceptualize the experimental data, a physical model was derived based on energetic considerations. Taken together, our analysis indicates that SV40, other polyoma viruses and some bacterial toxins (Shiga and cholera) use glycosphingolipids and a common pentameric protein scaffold to induce plasma membrane curvature, thus directly promoting their endocytic uptake into cells.

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Figure 1: SV40 infection and endocytosis depend on GM1 hydrocarbon chain structure.
Figure 2: SV40 binding induces caveolin-independent membrane invagination in cells.
Figure 3: SV40-induced membrane invagination on model membranes is dependent on GM1 hydrocarbon chain structure.
Figure 4: Induction of membrane invaginations by GM1-binding pentamer units.
Figure 5: Clustering and domain formation analysis.
Figure 6: Physical parameters controlling membrane invagination by SV40.

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Acknowledgements

This work was funded by the Swiss SystemsX.ch initiative, grant LipidX-2008/011 (to A.H.), the Human Frontier Science Program (to A.E.S., L.J. and P.S.), a FEBS fellowship (to H.E.), a CNRS fellowship (to W.R.), the Israel Science Foundation (Grant # 604/07 to A.O.), the Delegation Generale pour l'Armement (to L.B.), the Wellcome Trust (to W.C. and T.F.), Deutsche Forschungsgemeinschaft (to G.S.) and the European Regional Development Fund grant # 4212/04-01 (to K.B. and P.S.). H.E. thanks D. Choquet for his patience and support. The authors thank M. Abd-El-Latif for the preparation of VLPs and pentamers.

Author information

Author notes

  1. Helge Ewers

    Present address: Current address: ETH Zurich, Laboratory for Physical Chemistry, HCI F, Wolfgang-Pauli Strasse 10, 8093 Zurich, Switzerland.,

  2. Kirsten Bacia

    Present address: Current address: University of California at Berkeley, Department of Molecular and Cell Biology, Berkeley, CA 94720, USA.,

  3. Helge Ewers and Winfried Römer: These authors contributed equally to this work.

  4. Ari Helenius and Ludger Johannes: These authors contributed equally to this work.

Authors and Affiliations

  1. ETH Zurich, Institute of Biochemistry, HPM E, Schafmattstrasse 18, Zurich, 8093, Switzerland

    Helge Ewers, Alicia E. Smith, Roberta Mancini, Jürgen Kartenbeck & Ari Helenius

  2. Institut Curie, Centre de Recherche, Laboratoire Trafic, Signalisation et Ciblage Intracellulaires, Paris Cedex 05, 75248, France

    Winfried Römer, Valérie Chambon & Ludger Johannes

  3. CNRS UMR144, France

    Winfried Römer, Valérie Chambon & Ludger Johannes

  4. Institute for Biophysics, TU Dresden, BIOTEC, Tatzberg 47-51, Dresden, D-01307, Germany

    Kirsten Bacia & Petra Schwille

  5. UMR Gulliver CNRS-ESPCI 7083, 10 rue Vauquelin, Paris Cedex 05, 75231, France

    Serge Dmitrieff & Pierre Sens

  6. Glycosciences Laboratory, Imperial College London, Harrow, HA1 3UJ, Middlesex, UK

    Wengang Chai & Ten Feizi

  7. M010, German Cancer Research Center (DKFZ), Heidelberg, D-69120, Germany

    Jürgen Kartenbeck

  8. Institut Curie, Centre de Recherche, Laboratoire Physico-Chimie, Paris Cedex 05, 75248, France

    Ludwig Berland

  9. Université P. et M. Curie/CNRS UMR168, France

    Ludwig Berland

  10. Department of Hematology, The Hebrew University-Hadassah Medical School, Ein Kerem, 91120, Jerusalem, Israel

    Ariella Oppenheim

  11. LIMES, Membrane Biology and Lipid Biochemistry Unit c/o Kekulé-Institut für Organische Chemie und Biochemie der Universitaet Bonn, Bonn, 53123, Germany

    Günter Schwarzmann

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Contributions

H.E. and A.E.S. designed and performed the experiments involving infectious SV40; W.R. and V.C. designed and performed tubulation experiments in cells; A.E.S., J.K. and R.M. performed electron microscopy experiments; W.R., L.B., K.B. and Pe.S. designed and performed GUV experiments; W.C., G.S. and T.F. provided GM1 species; A.O. provided SV40 VLPs; S.D. and P.S. performed theoretical analysis; H.E., P.S., L.J. and A.H. wrote the manuscript; A.H. and L.J. supervised the work.

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Correspondence to Ari Helenius.

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Ewers, H., Römer, W., Smith, A. et al. GM1 structure determines SV40-induced membrane invagination and infection. Nat Cell Biol 12, 11–18 (2010). https://doi.org/10.1038/ncb1999

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