GM1 structure determines SV40-induced membrane invagination and infection


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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This work was funded by the Swiss 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.

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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).

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