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Live-cell visualization of dynamics of HIV budding site interactions with an ESCRT component

Nature Cell Biology volume 13, pages 469–474 (2011)Cite this article

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Abstract

HIV (human immunodeficiency virus) diverts the cellular ESCRT (endosomal sorting complex required for transport) machinery to promote virion release from infected cells. The ESCRT consists of four heteromeric complexes (ESCRT-0 to ESCRT-III), which mediate different membrane abscission processes, most importantly formation of intralumenal vesicles at multivesicular bodies. The ATPase VPS4 (vacuolar protein sorting 4) acts at a late stage of ESCRT function, providing energy for ESCRT dissociation. Recruitment of ESCRT by late-domain motifs in the viral Gag polyprotein and a role of ESCRT in HIV release are firmly established, but the order of events, their kinetics and the mechanism of action of individual ESCRT components in HIV budding are unclear at present. Using live-cell imaging, we show late-domain-dependent recruitment of VPS4A to nascent HIV particles at the host cell plasma membrane. Recruitment of VPS4A was transient, resulting in a single or a few bursts of at least two to five VPS4 dodecamers assembling at HIV budding sites. Bursts lasted for ∼35 s and appeared with variable delay before particle release. These results indicate that VPS4A has a direct role in membrane scission leading to HIV-1 release.

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Figure 1: Recruitment of VPS4A to HIV assembly sites.
Figure 2: Duration of eGFP–VPS4A bursts.
Figure 3: Number of eGFP–VPS4A molecules per burst.
Figure 4: Correlation of VPS4A wild-type bursts with Gag assembly phases.

Change history

  • 15 March 2011

    In the version of this letter initially published online, the first sentence was erroneously truncated and eGFP–VPS4A was misspelled in the second paragraph.

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Acknowledgements

We thank W. Sundquist (University of Utah, Salt Lake City) and R. Tsien (University of California, San Diego) for plasmids. We appreciate the assistance of M. Franke, M. Anders-Ößwein, A. M. Heuser, W. Schrimpf, D. Schupp and M. Eckhardt and tracking support from M. Preiner, F. Kohler and S. A. Rahman. We thank J. Hurley for helpful comments, A. Godin for Matlab masking algorithms and W. Godinez, J. P. Bergeestand, K. Rohr for support with the single-particle tracking algorithm. We gratefully acknowledge financial support of the DFG through SPP1175 (D.C.L., C.B., H-G.K.), SFB638 (H-G.K.) and grant MU885/4-2 (B.M.), the German Excellence Initiative through ‘Nanosystems Initiative Munich (NIM)’ (D.C.L., C.B.) and ‘CellNetworks’ (H-G.K.), the Ludwig-Maximilians-University Munich (LMUInnovativ BioImaging Network, D.C.L., C.B.), the EU (HIV-ACE network HEALTH-F3-2008-201095; B.M., H-G.K.), the Natural Sciences and Engineering Research Council of Canada (P.W.W.) and the Canadian Institutes of Health Research (P.W.W.).

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Authors and Affiliations

  1. Department of Chemistry and Biochemistry, Physical Chemistry, Munich Center for Integrated Protein Science (CiPSM) and Center for NanoScience, Ludwig-Maximilians-Universität München, Butenandtstraße 11, 81377 Munich, Germany

    Viola Baumgärtel, Sergey Ivanchenko, Aurélie Dupont, Christoph Bräuchle & Don C. Lamb

  2. Physics Department, McGill University, Montreal, Quebec H3A 2T8, Canada

    Mikhail Sergeev & Paul W. Wiseman

  3. Department of Infectious Diseases, Virology, Universitätsklinikum Heidelberg, Im Neuenheimer Feld 324, 69120 Heidelberg, Germany

    Hans-Georg Kräusslich & Barbara Müller

  4. Department of Physics, University of Illinois at Urbana-Champaign, Urbana, Illinois 61801, USA

    Don C. Lamb

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  1. Viola Baumgärtel
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Contributions

V.B. and S.I. carried out experiments. M.S. and P.W.W. carried out the ICS analysis. V.B., S.I. and A.D. analysed data. B.M., D.C.L., H-G.K., S.I., A.D., M.S., P.W.W., C.B. and V.B. wrote the manuscript. B.M., D.C.L., H-G.K., C.B. designed and guided the project.

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Correspondence to Barbara Müller or Don C. Lamb.

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The authors declare no competing financial interests.

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Baumgärtel, V., Ivanchenko, S., Dupont, A. et al. Live-cell visualization of dynamics of HIV budding site interactions with an ESCRT component. Nat Cell Biol 13, 469–474 (2011). https://doi.org/10.1038/ncb2215

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