Hafer Terry L, Felton Abby, Delgado Yennifer, Srinivasan Harini, Emerman Michael
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.
bioRxiv. 2023 Jul 28:2023.07.28.551016. doi: 10.1101/2023.07.28.551016.
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 , , , , , and . Knockout of Cyclin T1 ( ), a component of the P-TEFb complex 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 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 redundant in T cells but is absolutely required for HIV latency reversal.
HIV-1复制所需的宿主因子在潜伏期逆转中也发挥作用。利用一个包含假定HIV依赖因子的CRISPR基因文库，我们进行了一项筛选以鉴定潜伏期重新激活所需的基因。我们鉴定出了几个在HIV-1潜伏期重新激活中起关键作用的HIV-1依赖因子，包括 、 、 、 、 和 。细胞周期蛋白T1（ ）是对转录延伸很重要的P-TEFb复合物的一个组分，其敲除是筛选中的头号命中结果，并且对多种潜伏期逆转剂的HIV潜伏期逆转有最大影响。此外， 敲除可防止在HIV潜伏期的原代CD4+ T细胞模型中潜伏期重新激活，而不影响这些细胞的活化。RNA测序数据表明，CCNT1对HIV-1前病毒基因的调控程度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转录本没有显著影响。我们得出结论，CCNT1功能在T细胞中是冗余的，但对于HIV潜伏期逆转是绝对必需的。
