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HIV gp41–mediated membrane fusion occurs at edges of cholesterol-rich lipid domains

Nature Chemical Biology volume 11, pages 424–431 (2015)Cite this article

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Abstract

Lipid rafts in plasma membranes have emerged as possible platforms for the entry of HIV and other viruses into cells. However, little is known about how lipid phase heterogeneity contributes to viral entry because of the fine-grained and still poorly understood complexity of biological membranes. We used model systems mimicking HIV envelopes and T cell membranes and found that raft-like liquid-ordered (Lo-phase) lipid domains were necessary and sufficient for efficient membrane targeting and fusion. Interestingly, membrane binding and fusion were low in homogeneous liquid-disordered (Ld-phase) and Lo-phase membranes, indicating that lipid phase heterogeneity is essential. The HIV fusion peptide preferentially targeted to Lo-Ld boundary regions and promoted full fusion at the interface between ordered and disordered lipids. Ld-phase vesicles proceeded only to hemifusion. Thus, we propose that edges but not areas of raft-like ordered lipid domains are vital for HIV entry and membrane fusion.

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Figure 1: Phase-separated domains and cholesterol are required for HIV FP to fuse HIV envelope– and T cell membrane–mimicking lipid mixtures.
Figure 2: Membrane-bound HIV FP preferentially binds phase-separated Lo+d liposomes and targets Lo-Ld phase boundaries in phase-separated membranes.
Figure 3: HIV FP liposomes are targeted to Lo-Ld phase boundary regions in GUVs.
Figure 4: HIV FP liposomes fuse at Lo-Ld phase boundaries in supported membranes.
Figure 5: HIV-1 pseudoviruses bind preferentially to Lo-Ld phase boundaries in supported lipid bilayers or GUVs composed of bSM-DOPC-Ch (2:2:1).
Figure 6: Lipid domain boundaries in viral and cellular target membranes promote HIV gp41–mediated membrane fusion.

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Acknowledgements

This work was supported by NIH grants R01 AI30577 (to L.K.T.) and R21 AI103601 (to J.M.W.). We thank E. Nelson for technical help with the production of pseudoviruses.

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Authors and Affiliations

  1. Department of Molecular Physiology and Biological Physics, University of Virginia, Charlottesville, Virginia, USA

    Sung-Tae Yang, Volker Kiessling & Lukas K Tamm

  2. Center for Membrane Biology, University of Virginia, Charlottesville, Virginia, USA

    Sung-Tae Yang, Volker Kiessling, James A Simmons, Judith M White & Lukas K Tamm

  3. Department of Cell Biology, University of Virginia, Charlottesville, Virginia, USA

    James A Simmons & Judith M White

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Contributions

S.-T.Y., V.K., J.M.W. and L.K.T. designed research; S.-T.Y. performed most experiments; J.A.S. provided pseudotyped viruses; S.-T.Y., V.K., J.M.W. and L.K.T. analyzed data; and S.-T.Y., V.K. and L.K.T. wrote the paper.

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Correspondence to Lukas K Tamm.

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Supplementary information

Supplementary Text and Figures

Supplementary Results, Supplementary Figures 1–8 and Supplementary Table 1. (PDF 10999 kb)

Binding of LUVs to a GUV imaged by epifluorescence microscopy. (AVI 1389 kb)

Fusion of LUVs to a supported bilayer observed by TIRF microscopy. (AVI 9111 kb)

41589_2015_BFnchembio1800_MOESM444_ESM.avi

Fusion of a single liposome to a supported bilayer monitored by simultaneous observation of lipid (DiD) mixing (left panel) and content (sulforhodamine) release (center panel) and the DiD and sulforhodamine channels combined (right panel). (AVI 803 kb)

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Yang, ST., Kiessling, V., Simmons, J. et al. HIV gp41–mediated membrane fusion occurs at edges of cholesterol-rich lipid domains. Nat Chem Biol 11, 424–431 (2015). https://doi.org/10.1038/nchembio.1800

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