Structural alteration of DNA induced by viral protein R of HIV-1 triggers the DNA damage response
Abstract 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 o...
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| Format: | Article |
| Language: | English |
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BMC
2018-01-01
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| Series: | Retrovirology |
| Online Access: | http://link.springer.com/article/10.1186/s12977-018-0391-8 |
| _version_ | 1811263176906375168 |
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| author | Kenta Iijima Junya Kobayashi Yukihito Ishizaka |
| author_facet | Kenta Iijima Junya Kobayashi Yukihito Ishizaka |
| author_sort | Kenta Iijima |
| collection | DOAJ |
| description | Abstract 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. |
| first_indexed | 2024-04-12T19:39:59Z |
| format | Article |
| id | doaj.art-b785e0d6d444448592263dac665ffc26 |
| institution | Directory Open Access Journal |
| issn | 1742-4690 |
| language | English |
| last_indexed | 2024-04-12T19:39:59Z |
| publishDate | 2018-01-01 |
| publisher | BMC |
| record_format | Article |
| series | Retrovirology |
| spelling | doaj.art-b785e0d6d444448592263dac665ffc262022-12-22T03:19:07ZengBMCRetrovirology1742-46902018-01-0115112510.1186/s12977-018-0391-8Structural alteration of DNA induced by viral protein R of HIV-1 triggers the DNA damage responseKenta Iijima0Junya Kobayashi1Yukihito Ishizaka2Department of Intractable Diseases, National Center for Global Health and MedicineDepartment of Genome Repair Dynamics, Radiation Biology Center, Kyoto UniversityDepartment of Intractable Diseases, National Center for Global Health and MedicineAbstract 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.http://link.springer.com/article/10.1186/s12977-018-0391-8 |
| spellingShingle | Kenta Iijima Junya Kobayashi Yukihito Ishizaka Structural alteration of DNA induced by viral protein R of HIV-1 triggers the DNA damage response Retrovirology |
| 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 |
| url | http://link.springer.com/article/10.1186/s12977-018-0391-8 |
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