1. Cell Biology
  2. Microbiology and Infectious Disease
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HIV-1 uncoating by release of viral cDNA from capsid-like structures in the nucleus of infected cells

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Cite this article as: eLife 2021;10:e64776 doi: 10.7554/eLife.64776

Abstract

HIV-1 replication commences inside the cone-shaped viral capsid, but timing, localization and mechanism of uncoating are under debate. We adapted a strategy to visualize individual reverse-transcribed HIV-1 cDNA molecules and their association with viral and cellular proteins using fluorescence and correlative-light-and-electron-microscopy (CLEM). We specifically detected HIV-1 cDNA inside nuclei, but not in the cytoplasm. Nuclear cDNA initially co-localized with a fluorescent integrase fusion (IN-FP) and the viral CA (capsid) protein, but cDNA-punctae separated from IN-FP/CA over time. This phenotype was conserved in primary HIV-1 target cells, with nuclear HIV-1 complexes exhibiting strong CA-signals in all cell types. CLEM revealed cone-shaped HIV-1 capsid-like structures and apparently broken capsid-remnants at the position of IN-FP signals and elongated chromatin-like structures in the position of viral cDNA punctae lacking IN-FP. Our data argue for nuclear uncoating by physical disruption rather than cooperative disassembly of the CA-lattice, followed by physical separation from the pre-integration complex.

Data availability

All data generated or analysed during this study are included in the manuscript and supporting files. Source data files have been provided for Figure 1f,h, Figure 1 figure supplement 1c-d, Figure 1 figure supplement 3a, Figure 2a-b, Figure 2e-f, Figure 3b-c, Figure 4c,g,h,j-k,o, Figure 4 - figure supplement 1a-b, Figure 7c,e-f, Figure 7 - figure supplement 1b, Figure 7 - figure supplement 2a-b. Materials involving the ANCHOR system are MTA-restricted and commercially available from NeoVirTech (France).

Article and author information

Author details

  1. Thorsten G Müller

    Department of Infectious Diseases, Virology, University Hospital Heidelberg, Heidelberg, Germany
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0002-4197-6224
  2. Vojtech Zila

    Department of Infectious Diseases, Virology, University Hospital Heidelberg, Heidelberg, Germany
    Competing interests
    The authors declare that no competing interests exist.
  3. Kyra Peters

    Department of Infectious Diseases, Virology, University Hospital Heidelberg, Heidelberg, Germany
    Competing interests
    The authors declare that no competing interests exist.
  4. Sandra Schifferdecker

    Department of Infectious Diseases, Virology, University Hospital Heidelberg, Heidelberg, Germany
    Competing interests
    The authors declare that no competing interests exist.
  5. Mia Stanic

    Department of Infectious Diseases, Integrative Virology, University Hospital Heidelberg, Heidelberg, Germany
    Competing interests
    The authors declare that no competing interests exist.
  6. Bojana Lucic

    Center for Integrative Infectious Diseases, University Hospital Heidelberg, Heidelberg, Germany
    Competing interests
    The authors declare that no competing interests exist.
  7. Vibor Laketa

    Department of Infectious Diseases, Virology, University Hospital Heidelberg, Heidelberg, Germany
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0002-9472-2738
  8. Marina Lusic

    Center for Integrative Infectious Diseases, University Hospital Heidelberg, Heidelberg, Germany
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0002-0120-3569
  9. Barbara Müller

    Department of Infectious Diseases, Virology, University Hospital Heidelberg, Heidelberg, Germany
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0001-5726-5585
  10. Hans-Georg Kräusslich

    Department of Infectious Diseases, Virology, University Hospital Heidelberg, Heidelberg, Germany
    For correspondence
    hans-georg.kraeusslich@med.uni-heidelberg.de
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0002-8756-329X

Funding

Deutsche Forschungsgemeinschaft (Projektnummer 240245660 - SFB 1129 project 5)

  • Thorsten G Müller
  • Vojtech Zila
  • Kyra Peters
  • Hans-Georg Kräusslich

Deutsche Forschungsgemeinschaft (Projektnummer 240245660 - SFB 1129 project 6)

  • Sandra Schifferdecker
  • Barbara Müller

Deutsche Forschungsgemeinschaft (Projektnummer 240245660 - SFB 1129 project 20)

  • Mia Stanic
  • Bojana Lucic
  • Marina Lusic

Deutsches Zentrum für Infektionsforschung (TTU HIV)

  • Vibor Laketa
  • Marina Lusic
  • Hans-Georg Kräusslich

The funders had no role in study design, data collection and interpretation, or the decision to submit the work for publication.

Reviewing Editor

  1. Wesley I Sundquist, University of Utah School of Medicine, United States

Publication history

  1. Received: November 10, 2020
  2. Accepted: April 21, 2021
  3. Accepted Manuscript published: April 27, 2021 (version 1)
  4. Version of Record published: June 1, 2021 (version 2)

Copyright

© 2021, Müller et al.

This article is distributed under the terms of the Creative Commons Attribution License permitting unrestricted use and redistribution provided that the original author and source are credited.

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