Role of ATM and DNA‐PKcs in early replicative stages of HIV‐1 life cycle

Anisenko, A.; Nefyodova, A.; Gottikh, M.

Авторы: Anisenko A., Nefyodova A., Gottikh M.
Журнал: Molecular Biology of the Cell
Том: 31
Номер: 26
Год издания: 2020
Издательство: American Society for Cell Biology
Местоположение издательства: United States
Номер статьи: P1122
DOI: 10.1091/mbc.E20-10-0665
Аннотация: HIV‐1 integration results in genomic DNA gaps that are repaired by cellular DNA repair pathways. Previously, we demonstrated participation of the nonhomologous end‐joining repair factors (NHEJ) such as Ku70, Ku80 and DNA‐PKcs in the post‐integrational gap repair (PIR), which is initiated by direct interaction of HIV‐1 integrase with Ku70. This interaction leads to DNA‐PKcs binding and subsequent activation. Inhibition of DNA‐PKcs by small inhibitor Nu7441 decreases the gap repair efficiency and as a result efficiency of cell transduction by HIV‐1 based pseudovirus. DNA‐PKcs belongs to phosphatidylinositol 3‐kinase‐related kinase family (PIKKs), including two other DNA damage response kinases: ATM and ATR. To test their contribution to post‐integrational repair, we first analyzed the effect of Ku‐55933 (ATM inhibitor), AZ20 (ATR inhibitor) and Nu7441 (DNA‐PKcs inhibitor, as a positive control) on the 293T transduction efficiency by VSV‐G‐pseudotyped HIV‐1 like particles. Inhibition of ATM or DNA‐PKcs but not ATR kinase activity decreased luciferase production (Luc). Interestingly, inhibitors of ATM and DNA‐PKcs didn’t affect Luc production in case of pseudovirus with mutant integrase defective in Ku binding and subsequently in PIR. The qPCR data revealed the effect of ATM or DNA‐PKcs inhibition on post‐integrational DNA repair, but not on reverse transcription or integration. Kinetics of DNA‐PKcs and ATM activation were further studied using time of drug addition assay. We found that activation of these kinases occurs simultaneously with integration of viral cDNA (7‐14 hpi). Taking into account the fact that the first repaired forms appear with a significant time lag after integration (16‐18 hpi), it can be assumed that DNA‐PKcs and ATM just initiate PIR, and further repair processes require additional factors. This work was supported by RSF grant 19‐74‐10021.
Добавил в систему: Анисенко Андрей Николаевич

	ИСТИНА	Войти в систему Регистрация
	ФНКЦ РР
	Главная Поиск Статистика О проекте Помощь

ИСТИНА

ФНКЦ РР

Role of ATM and DNA‐PKcs in early replicative stages of HIV‐1 life cycleстатья Тезисы

Прикрепленные файлы