HIV-1 capsid is involved in post-nuclear entry steps

Nan Yu Chen, Lihong Zhou, Paul J. Gane, Silvana Opp, Neil J. Ball, Giuseppe Nicastro, Madeleine Zufferey, Cindy Buffone, Jeremy Luban, David Selwood, Felipe Diaz-Griffero, Ian Taylor, Ariberto Fassati

Research output: Contribution to journalArticle

20 Citations (Scopus)

Abstract

Background: HIV-1 capsid influences viral uncoating and nuclear import. Some capsid is detected in the nucleus but it is unclear if it has any function. We reported that the antibiotic Coumermycin-A1 (C-A1) inhibits HIV-1 integration and that a capsid mutation confers resistance to C-A1, suggesting that capsid might affect post-nuclear entry steps. Results: Here we report that C-A1 inhibits HIV-1 integration in a capsid-dependent way. Using molecular docking, we identify an extended binding pocket delimited by two adjacent capsid monomers where C-A1 is predicted to bind. Isothermal titration calorimetry confirmed that C-A1 binds to hexameric capsid. Cyclosporine washout assays in Jurkat CD4+ T cells expressing engineered human TRIMCyp showed that C-A1 causes faster and greater escape from TRIMCyp restriction. Sub-cellular fractionation showed that small amounts of capsid accumulated in the nuclei of infected cells and C-A1 reduced the nuclear capsid. A105S and N74D capsid mutant viruses did not accumulate capsid in the nucleus, irrespective of C-A1 treatment. Depletion of Nup153, a nucleoporin located at the nuclear side of the nuclear pore that binds to HIV-1 capsid, made the virus less susceptible to TRIMCyp restriction, suggesting that Nup153 may help maintain some integrity of the viral core in the nucleus. Furthermore C-A1 increased binding of CPSF6, a nuclear protein, to capsid. Conclusions: Our results indicate that capsid is involved in post-nuclear entry steps preceding integration.

Original languageEnglish (US)
Article number28
JournalRetrovirology
Volume13
Issue number1
DOIs
StatePublished - 2016

Fingerprint

Capsid
HIV-1
Virus Uncoating
Nuclear Pore Complex Proteins
Viruses
Nuclear Pore
coumermycin
Calorimetry
Cell Nucleus Active Transport
Nuclear Proteins
Cell Nucleus
Cyclosporine

Keywords

  • Capsid
  • Coumermycin-A1
  • HIV-1
  • Integration
  • Nucleoporins
  • Nucleus
  • Nup153
  • Uncoating

ASJC Scopus subject areas

  • Infectious Diseases
  • Virology

Cite this

Chen, N. Y., Zhou, L., Gane, P. J., Opp, S., Ball, N. J., Nicastro, G., ... Fassati, A. (2016). HIV-1 capsid is involved in post-nuclear entry steps. Retrovirology, 13(1), [28]. https://doi.org/10.1186/s12977-016-0262-0

HIV-1 capsid is involved in post-nuclear entry steps. / Chen, Nan Yu; Zhou, Lihong; Gane, Paul J.; Opp, Silvana; Ball, Neil J.; Nicastro, Giuseppe; Zufferey, Madeleine; Buffone, Cindy; Luban, Jeremy; Selwood, David; Diaz-Griffero, Felipe; Taylor, Ian; Fassati, Ariberto.

In: Retrovirology, Vol. 13, No. 1, 28, 2016.

Research output: Contribution to journalArticle

Chen, NY, Zhou, L, Gane, PJ, Opp, S, Ball, NJ, Nicastro, G, Zufferey, M, Buffone, C, Luban, J, Selwood, D, Diaz-Griffero, F, Taylor, I & Fassati, A 2016, 'HIV-1 capsid is involved in post-nuclear entry steps', Retrovirology, vol. 13, no. 1, 28. https://doi.org/10.1186/s12977-016-0262-0
Chen NY, Zhou L, Gane PJ, Opp S, Ball NJ, Nicastro G et al. HIV-1 capsid is involved in post-nuclear entry steps. Retrovirology. 2016;13(1). 28. https://doi.org/10.1186/s12977-016-0262-0
Chen, Nan Yu ; Zhou, Lihong ; Gane, Paul J. ; Opp, Silvana ; Ball, Neil J. ; Nicastro, Giuseppe ; Zufferey, Madeleine ; Buffone, Cindy ; Luban, Jeremy ; Selwood, David ; Diaz-Griffero, Felipe ; Taylor, Ian ; Fassati, Ariberto. / HIV-1 capsid is involved in post-nuclear entry steps. In: Retrovirology. 2016 ; Vol. 13, No. 1.
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abstract = "Background: HIV-1 capsid influences viral uncoating and nuclear import. Some capsid is detected in the nucleus but it is unclear if it has any function. We reported that the antibiotic Coumermycin-A1 (C-A1) inhibits HIV-1 integration and that a capsid mutation confers resistance to C-A1, suggesting that capsid might affect post-nuclear entry steps. Results: Here we report that C-A1 inhibits HIV-1 integration in a capsid-dependent way. Using molecular docking, we identify an extended binding pocket delimited by two adjacent capsid monomers where C-A1 is predicted to bind. Isothermal titration calorimetry confirmed that C-A1 binds to hexameric capsid. Cyclosporine washout assays in Jurkat CD4+ T cells expressing engineered human TRIMCyp showed that C-A1 causes faster and greater escape from TRIMCyp restriction. Sub-cellular fractionation showed that small amounts of capsid accumulated in the nuclei of infected cells and C-A1 reduced the nuclear capsid. A105S and N74D capsid mutant viruses did not accumulate capsid in the nucleus, irrespective of C-A1 treatment. Depletion of Nup153, a nucleoporin located at the nuclear side of the nuclear pore that binds to HIV-1 capsid, made the virus less susceptible to TRIMCyp restriction, suggesting that Nup153 may help maintain some integrity of the viral core in the nucleus. Furthermore C-A1 increased binding of CPSF6, a nuclear protein, to capsid. Conclusions: Our results indicate that capsid is involved in post-nuclear entry steps preceding integration.",
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AU - Zhou, Lihong

AU - Gane, Paul J.

AU - Opp, Silvana

AU - Ball, Neil J.

AU - Nicastro, Giuseppe

AU - Zufferey, Madeleine

AU - Buffone, Cindy

AU - Luban, Jeremy

AU - Selwood, David

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AU - Taylor, Ian

AU - Fassati, Ariberto

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AB - Background: HIV-1 capsid influences viral uncoating and nuclear import. Some capsid is detected in the nucleus but it is unclear if it has any function. We reported that the antibiotic Coumermycin-A1 (C-A1) inhibits HIV-1 integration and that a capsid mutation confers resistance to C-A1, suggesting that capsid might affect post-nuclear entry steps. Results: Here we report that C-A1 inhibits HIV-1 integration in a capsid-dependent way. Using molecular docking, we identify an extended binding pocket delimited by two adjacent capsid monomers where C-A1 is predicted to bind. Isothermal titration calorimetry confirmed that C-A1 binds to hexameric capsid. Cyclosporine washout assays in Jurkat CD4+ T cells expressing engineered human TRIMCyp showed that C-A1 causes faster and greater escape from TRIMCyp restriction. Sub-cellular fractionation showed that small amounts of capsid accumulated in the nuclei of infected cells and C-A1 reduced the nuclear capsid. A105S and N74D capsid mutant viruses did not accumulate capsid in the nucleus, irrespective of C-A1 treatment. Depletion of Nup153, a nucleoporin located at the nuclear side of the nuclear pore that binds to HIV-1 capsid, made the virus less susceptible to TRIMCyp restriction, suggesting that Nup153 may help maintain some integrity of the viral core in the nucleus. Furthermore C-A1 increased binding of CPSF6, a nuclear protein, to capsid. Conclusions: Our results indicate that capsid is involved in post-nuclear entry steps preceding integration.

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