The Role of DNA Repair Complex DNA-PK in HIV-1 Transcription

Shadrina, O.; Anisenko, A.; Gottikh, M.

Авторы: Shadrina Olga, Anisenko Andrey, Gottikh Marina
Сборник: Proceedings
Том: 50
Тезисы
Год издания: 2020
Место издания: MDPI AG
Первая страница: 133
Последняя страница: 133
DOI: 10.3390/proceedings2020050133
Аннотация: The human DNA-dependent protein kinase (DNA-PK), composed of the heterodimericprotein Ku and catalytic subunit DNA-PKcs, is a sensor of double-strand DNA breaks in the nonhomologous end-joining DNA repair pathway. The key role of DNA-PK in the post-integrationalrepair of HIV-1 has been shown. It has also been suggested that DNA-PK can participate in theregulation of HIV transcription, although the mechanism is unclear. To clarify the impact of eachDNA-PK subunit on the transcription of HIV-1, HEK 293T cells, in which each of the DNA-PKcomponents was depleted, were transfected with reporter vectors containing firefly luciferase underthe control of HIV LTR promoter. We detected a positive influence of both Ku subunits, but not ofDNA-PKcs, on the transcription from the HIV promoter. Ku is known to interact with HIV-1 TARRNA, playing an essential role in viral transcription; nonetheless, the deletion of the TAR-codingregion from LTR did not alter the Ku effect. Human small noncoding 7SK RNA participates in HIV1 transcription. The direct binding of recombinant Ku and in vitro transcribed 7SK RNA wasdemonstrated using EMSA. In addition, we identified the interactions of endogenous Ku withproteins HEXIM1 and Cdk9 from the 7SK RNP complex. These results suggest that Ku exerts itseffects on HIV-1 transcription via interaction with the 7SK RNP complex. However, we cannot ruleout an indirect effect of Ku on transcription via the regulation of the levels of some transcriptionfactors participating in HIV-1 transcription. We performed a transcriptome analysis of wild typeHEK 293T cells and those with depleted DNA-PK subunits. The genes regulated by each subunitwere defined and the genes that were mainly dependent on Ku subunits were selected. Amongthem, we identified transcription factors enhancing HIV-1 transcription, whose levels weredownregulated in Ku-depleted cells. The study was supported by RFBR grant №18-04-00542 andRSF grant №17-14-01107.
Добавил в систему: Анисенко Андрей Николаевич

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