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Structural alteration of DNA induced by viral protein R of HIV-1 triggers the DNA damage response
BACKGROUND: Viral protein R (Vpr) is an accessory protein of HIV-1, which is potentially involved in the infection of macrophages and the induction of the ataxia-telangiectasia and Rad3-related protein (ATR)-mediated DNA damage response (DDR). It was recently proposed that the SLX4 complex of struct...
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Formato: | Online Artículo Texto |
Lenguaje: | English |
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BioMed Central
2018
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5771197/ https://www.ncbi.nlm.nih.gov/pubmed/29338752 http://dx.doi.org/10.1186/s12977-018-0391-8 |
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author | Iijima, Kenta Kobayashi, Junya Ishizaka, Yukihito |
author_facet | Iijima, Kenta Kobayashi, Junya Ishizaka, Yukihito |
author_sort | Iijima, Kenta |
collection | PubMed |
description | BACKGROUND: Viral protein R (Vpr) is an accessory protein of HIV-1, which is potentially involved in the infection of macrophages and the induction of the ataxia-telangiectasia and Rad3-related protein (ATR)-mediated DNA damage response (DDR). It was recently proposed that the SLX4 complex of structure-specific endonuclease is involved in Vpr-induced DDR, which implies that aberrant DNA structures are responsible for this phenomenon. However, the mechanism by which Vpr alters the DNA structures remains unclear. RESULTS: We found that Vpr unwinds double-stranded DNA (dsDNA) and invokes the loading of RPA70, which is a single-stranded DNA-binding subunit of RPA that activates the ATR-dependent DDR. We demonstrated that Vpr influenced RPA70 to accumulate in the corresponding region utilizing the LacO/LacR system, in which Vpr can be tethered to the LacO locus. Interestingly, RPA70 recruitment required chromatin remodelling via Vpr-mediated ubiquitination of histone H2B. On the contrary, Q65R mutant of Vpr, which lacks ubiquitination activity, was deficient in both chromatin remodelling and RPA70 loading on to the chromatin. Moreover, Vpr-induced unwinding of dsDNA coincidently resulted in the accumulation of negatively supercoiled DNA and covalent complexes of topoisomerase 1 and DNA, which caused DNA double-strand breaks (DSBs) and DSB-directed integration of proviral DNA. Lastly, we noted the dependence of Vpr-promoted HIV-1 infection in resting macrophages on topoisomerase 1. CONCLUSIONS: The findings of this study indicate that Vpr-induced structural alteration of DNA is a primary event that triggers both DDR and DSB, which ultimately contributes to HIV-1 infection. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (10.1186/s12977-018-0391-8) contains supplementary material, which is available to authorized users. |
format | Online Article Text |
id | pubmed-5771197 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2018 |
publisher | BioMed Central |
record_format | MEDLINE/PubMed |
spelling | pubmed-57711972018-01-25 Structural alteration of DNA induced by viral protein R of HIV-1 triggers the DNA damage response Iijima, Kenta Kobayashi, Junya Ishizaka, Yukihito Retrovirology Research BACKGROUND: Viral protein R (Vpr) is an accessory protein of HIV-1, which is potentially involved in the infection of macrophages and the induction of the ataxia-telangiectasia and Rad3-related protein (ATR)-mediated DNA damage response (DDR). It was recently proposed that the SLX4 complex of structure-specific endonuclease is involved in Vpr-induced DDR, which implies that aberrant DNA structures are responsible for this phenomenon. However, the mechanism by which Vpr alters the DNA structures remains unclear. RESULTS: We found that Vpr unwinds double-stranded DNA (dsDNA) and invokes the loading of RPA70, which is a single-stranded DNA-binding subunit of RPA that activates the ATR-dependent DDR. We demonstrated that Vpr influenced RPA70 to accumulate in the corresponding region utilizing the LacO/LacR system, in which Vpr can be tethered to the LacO locus. Interestingly, RPA70 recruitment required chromatin remodelling via Vpr-mediated ubiquitination of histone H2B. On the contrary, Q65R mutant of Vpr, which lacks ubiquitination activity, was deficient in both chromatin remodelling and RPA70 loading on to the chromatin. Moreover, Vpr-induced unwinding of dsDNA coincidently resulted in the accumulation of negatively supercoiled DNA and covalent complexes of topoisomerase 1 and DNA, which caused DNA double-strand breaks (DSBs) and DSB-directed integration of proviral DNA. Lastly, we noted the dependence of Vpr-promoted HIV-1 infection in resting macrophages on topoisomerase 1. CONCLUSIONS: The findings of this study indicate that Vpr-induced structural alteration of DNA is a primary event that triggers both DDR and DSB, which ultimately contributes to HIV-1 infection. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (10.1186/s12977-018-0391-8) contains supplementary material, which is available to authorized users. BioMed Central 2018-01-16 /pmc/articles/PMC5771197/ /pubmed/29338752 http://dx.doi.org/10.1186/s12977-018-0391-8 Text en © The Author(s) 2018 Open AccessThis article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated. |
spellingShingle | Research Iijima, Kenta Kobayashi, Junya Ishizaka, Yukihito Structural alteration of DNA induced by viral protein R of HIV-1 triggers the DNA damage response |
title | Structural alteration of DNA induced by viral protein R of HIV-1 triggers the DNA damage response |
title_full | Structural alteration of DNA induced by viral protein R of HIV-1 triggers the DNA damage response |
title_fullStr | Structural alteration of DNA induced by viral protein R of HIV-1 triggers the DNA damage response |
title_full_unstemmed | Structural alteration of DNA induced by viral protein R of HIV-1 triggers the DNA damage response |
title_short | Structural alteration of DNA induced by viral protein R of HIV-1 triggers the DNA damage response |
title_sort | structural alteration of dna induced by viral protein r of hiv-1 triggers the dna damage response |
topic | Research |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5771197/ https://www.ncbi.nlm.nih.gov/pubmed/29338752 http://dx.doi.org/10.1186/s12977-018-0391-8 |
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