HIV-1 nuclear import in macrophages is regulated by CPSF6-capsid interactions at the Nuclear Pore Complex

Abstract

Nuclear entry of HIV-1 replication complexes through intact nuclear pore complexes is critical for successful infection. The host protein cleavage-and-polyadenylation-specificity-factor-6 (CPSF6) has been implicated in different stages of early HIV-1 replication. Applying quantitative microscopy of HIV-1 reverse-transcription and pre-integration-complexes (RTC/PIC), we show that CPSF6 is strongly recruited to nuclear replication complexes but absent from cytoplasmic RTC/PIC in primary human macrophages. Depletion of CPSF6 or lack of CPSF6 binding led to accumulation of HIV-1 subviral complexes at the nuclear envelope of macrophages and reduced infectivity. Two-color stimulated-emission-depletion microscopy indicated that under these circumstances HIV-1 complexes are retained inside the nuclear pore and undergo CA-multimer dependent CPSF6 clustering adjacent to the nuclear basket. We propose that nuclear entry of HIV-1 subviral complexes in macrophages is mediated by consecutive binding of Nup153 and CPSF6 to the hexameric CA lattice.

Data availability

All data generated or analysed during this study are included in the manuscript and supporting files. Source data files for the plots of Figures 1,3,4 and supplemental material are provided.

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Author details

  1. David Alejandro Bejarano

    Department of Infectious Diseases, Virology, University of Heidelberg, Heidelberg, Germany
    Competing interests
    The authors declare that no competing interests exist.
  2. Ke Peng

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

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

    Department of Infectious Diseases, Virology, University of Heidelberg, Heidelberg, Germany
    Competing interests
    The authors declare that no competing interests exist.
  5. K Laurence Jost

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

    Center for Integrative Infectious Diseases, University of Heidelberg, Heidelberg, Germany
    Competing interests
    The authors declare that no competing interests exist.
  7. Bärbel Glass

    Department of Infectious Diseases, Virology, University of Heidelberg, Heidelberg, Germany
    Competing interests
    The authors declare that no competing interests exist.
  8. Marina Lusic

    Center for Integrative Infectious Diseases, University of 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 of 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 of 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 (SFB1129)

  • Barbara Müller
  • Hans-Georg Kräusslich

Deutsches Zentrum für Infektionsforschung (TTU HIV)

  • Hans-Georg Kräusslich

Deutsche Forschungsgemeinschaft (SPP1923)

  • Hans-Georg Kräusslich

Heidelberg Biosciences International Graduate School

  • David Alejandro Bejarano

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: September 7, 2018
  2. Accepted: January 16, 2019
  3. Accepted Manuscript published: January 23, 2019 (version 1)
  4. Version of Record published: March 5, 2019 (version 2)

Copyright

© 2019, Bejarano 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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  1. David Alejandro Bejarano
  2. Ke Peng
  3. Vibor Laketa
  4. Kathleen Börner
  5. K Laurence Jost
  6. Bojana Lucic
  7. Bärbel Glass
  8. Marina Lusic
  9. Barbara Müller
  10. Hans-Georg Kräusslich
(2019)
HIV-1 nuclear import in macrophages is regulated by CPSF6-capsid interactions at the Nuclear Pore Complex
eLife 8:e41800.
https://doi.org/10.7554/eLife.41800
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