Molecular and Cellular Biology Graduate Program, University of Washington, Seattle, WA 98195, USA.
Divisions of Human Biology and Basic Sciences, Fred Hutchinson Cancer Center, Seattle, WA 98109, USA.
Viruses. 2023 Aug 31;15(9):1863. doi: 10.3390/v15091863.
We sought to explore the hypothesis that host factors required for HIV-1 replication also play a role in latency reversal. Using a CRISPR gene library of putative HIV dependency factors, we performed a screen to identify genes required for latency reactivation. We identified several HIV-1 dependency factors that play a key role in HIV-1 latency reactivation including ELL, UBE2M, TBL1XR1, HDAC3, AMBRA1, and ALYREF. The knockout of Cyclin T1 (), a component of the P-TEFb complex that is important for transcription elongation, was the top hit in the screen and had the largest effect on HIV latency reversal with a wide variety of latency reversal agents. Moreover, knockout prevents latency reactivation in a primary CD4+ T cell model of HIV latency without affecting the activation of these cells. RNA sequencing data showed that CCNT1 regulates HIV-1 proviral genes to a larger extent than any other host gene and had no significant effects on RNA transcripts in primary T cells after activation. We conclude that CCNT1 function is non-essential in T cells but is absolutely required for HIV latency reversal.
我们试图探索这样一个假设，即 HIV-1 复制所需的宿主因素也在潜伏期逆转中发挥作用。我们使用了一个可能的 HIV 依赖因子的 CRISPR 基因文库，进行了一项筛选，以鉴定出潜伏激活所需的基因。我们鉴定出了几个在 HIV-1 潜伏期逆转中起关键作用的 HIV 依赖因子，包括 ELL、UBE2M、TBL1XR1、HDAC3、AMBRA1 和 ALYREF。敲除周期蛋白 T1（）是 P-TEFb 复合物的一个组成部分，对转录延伸很重要，它是筛选中的头号命中靶点，并且对各种潜伏期逆转剂的 HIV 潜伏期逆转有最大的影响。此外，敲除阻止了 HIV 潜伏期的重新激活，而不会影响这些细胞的激活在 HIV 潜伏期的原代 CD4+T 细胞模型中。RNA 测序数据表明，CCNT1 对 HIV-1 前病毒基因的调节作用大于任何其他宿主基因，并且在激活后对原代 T 细胞中的 RNA 转录物没有显著影响。我们得出的结论是，CCNT1 功能在 T 细胞中不是必需的，但对于 HIV 潜伏期的逆转是绝对必需的。
