HIV-1 capsid is involved in post-nuclear entry steps
| Autor: | Nan Yu Chen, Madeleine Zufferey, Paul J. Gane, Neil J. Ball, Cindy Buffone, Lihong Zhou, Giuseppe Nicastro, David L. Selwood, Felipe Diaz-Griffero, Ian A. Taylor, Ariberto Fassati, Jeremy Luban, Silvana Opp |
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| Rok vydání: | 2016 |
| Předmět: |
Aminocoumarins
0301 basic medicine viruses HIV Core Protein p24 Integration Biology Uncoating Antiviral Agents Jurkat cells Virus Nucleus Cell Line 03 medical and health sciences Capsid Virology Humans Nup153 Nuclear pore Nuclear protein Research Capsomere Nucleoporins Virus Internalization biochemical phenomena metabolism and nutrition 3. Good health Cell biology 030104 developmental biology Infectious Diseases Coumermycin-A1 HIV-1 Nucleoporin Nuclear transport |
| Zdroj: | Retrovirology |
| Popis: | 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. Electronic supplementary material The online version of this article (doi:10.1186/s12977-016-0262-0) contains supplementary material, which is available to authorized users. |
| Databáze: | OpenAIRE |
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