Li Zichong, Deeks Steven G, Ott Melanie, Greene Warner C
Gladstone Institute of Virology, San Francisco, CA 94158, USA.
University of California, San Francisco, San Francisco, CA 94143, USA.
Cell Rep. 2023 Sep 26;42(9):113055. doi: 10.1016/j.celrep.2023.113055. Epub 2023 Sep 7.
Host repressors mediate HIV latency, but how they interactively silence the virus remains unclear. Here, we develop "reiterative enrichment and authentication of CRISPRi targets for synergies (REACTS)" to probe the genome for synergies between HIV transcription repressors. Using eight known host repressors as queries, we identify 32 synergies involving eleven repressors, including BCL7C, KANSL2, and SIRT2. Overexpression of these three proteins reduces HIV reactivation in Jurkat T cells and in CD4 T cells from people living with HIV on antiretroviral therapy (ART). We show that the BCL7C-containing BAF complex and the KANSL2-containing NSL complex form a "supercomplex" that increases inhibitory histone acetylation of the HIV long-terminal repeat (LTR) and its occupancy by the short variant of the acetyl-lysine reader Brd4. Collectively, we provide a validated platform for defining gene synergies genome wide, and the BAF-NSL "supercomplex" represents a potential target for overcoming HIV rebound after ART cessation.
宿主抑制因子介导HIV潜伏，但它们如何协同沉默病毒仍不清楚。在此，我们开发了“用于协同作用的CRISPRi靶点的反复富集和验证（REACTS）”技术，以在基因组中探寻HIV转录抑制因子之间的协同作用。以八个已知的宿主抑制因子作为查询对象，我们鉴定出32种涉及11种抑制因子的协同作用，包括BCL7C、KANSL2和SIRT2。这三种蛋白质的过表达降低了Jurkat T细胞以及接受抗逆转录病毒疗法（ART）的HIV感染者的CD4 T细胞中的HIV重新激活。我们发现，含有BCL7C的BAF复合物和含有KANSL2的NSL复合物形成了一种“超级复合物”，该复合物增加了HIV长末端重复序列（LTR）的抑制性组蛋白乙酰化及其被乙酰赖氨酸阅读器Brd4短变体的占据。总体而言，我们提供了一个经过验证的全基因组定义基因协同作用的平台，并且BAF-NSL“超级复合物”代表了克服ART停药后HIV反弹的潜在靶点。
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