Chemistry Department, Lomonosov Moscow State University, 119992 Moscow, Russia.
Faculty of Bioengineering and Bioinformatics, Lomonosov Moscow State University, 119992 Moscow, Russia.
Int J Mol Sci. 2023 Feb 1;24(3):2797. doi: 10.3390/ijms24032797.
The integration of a DNA copy of an HIV-1 RNA genome into the host genome, carried out by the viral enzyme integrase, results in the formation of single-stranded gaps in cellular DNA that must be repaired. Here, we have analyzed the involvement of the PI3K kinases, ATM, ATR, and DNA-PKcs, which are important players in the DNA damage response (DDR) in HIV-1 post-integrational DNA repair (PIR). The participation of the DNA-PK complex in HIV-1 PIR has been previously shown, and the formation of a complex between the viral integrase and the DNA-PK subunit, Ku70, has been found to be crucial for efficient PIR. Now, we have shown that the inhibition of both DNA-PKcs and ATM, but not ATR, significantly reduces PIR efficiency. The activation of both kinases is a sequential process, where one kinase, being activated, activates the other, and it occurs simultaneously with the integration of viral DNA. This fact suggests that the activation of both kinases triggers PIR. Most interestingly, the activation of not only DNA-PKcs, but also ATM depends on the complex formation between integrase and Ku70. The elucidation of the interactions between viruses and DDR is important both for understanding the modulation of host cell functions by these pathogens and for developing new approaches to combat viral infections.
HIV-1 RNA 基因组的 DNA 拷贝通过病毒酶整合酶整合到宿主基因组中,导致细胞 DNA 中形成单链缺口,必须进行修复。在这里,我们分析了 PI3K 激酶、ATM、ATR 和 DNA-PKcs 的参与,它们在 HIV-1 整合后 DNA 修复 (PIR) 中的 DNA 损伤反应 (DDR) 中是重要的参与者。已经证明 DNA-PK 复合物参与 HIV-1 PIR,并且发现病毒整合酶和 DNA-PK 亚基 Ku70 之间形成复合物对于有效的 PIR 至关重要。现在,我们已经表明,抑制 DNA-PKcs 和 ATM,但不是 ATR,可显著降低 PIR 效率。两种激酶的激活是一个连续的过程,其中一种激酶被激活,激活另一种激酶,并且它与病毒 DNA 的整合同时发生。这一事实表明,两种激酶的激活触发了 PIR。最有趣的是,不仅 DNA-PKcs 的激活,而且 ATM 的激活都依赖于整合酶和 Ku70 之间的复合物形成。阐明病毒与 DDR 之间的相互作用对于理解这些病原体对宿主细胞功能的调节以及开发对抗病毒感染的新方法都很重要。