A CRISPR/Cas9 screen identifies the histone demethylase MINA53 as a novel HIV-1 latency-promoting gene (LPG)
A CRISPR/Cas9 screen identifies the histone demethylase MINA53 as a novel HIV-1 latency-promoting gene (LPG)
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DOI：
10.1093/nar/gkz493
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发表时间：
2019-08-22
影响因子：
14.9
通讯作者：
Zhu, Jian
中科院分区：
文献类型：
--
作者：
Huang, Huachao;Kong, Weili;Zhu, Jian
虽然联合抗逆转录病毒疗法能够有效阻断艾滋病患者体内HIV-1的活跃复制，但HIV-1在感染细胞中仍以转录失活的前病毒形式存在。这些HIV-1潜伏库仍然是清除HIV-1的主要障碍。研究调节HIV-1潜伏期的宿主因素对于开发新型抗逆转录病毒药物以消除HIV-1潜伏库至关重要。从我们最近完成的CRISPR/Cas9 sgRNA筛选中，我们确定了组蛋白去甲基化酶MINA 53可能是一种新的HIV-1潜伏促进基因（LPG）。我们接下来通过使用替代RNAi方法耗尽MINA 53来验证MINA 53在维持HIV-1潜伏期中的功能。我们进一步鉴定了体外MINA 53优先使组蛋白底物H3 K36 me 3去甲基化，并且在细胞中通过RNAi消耗MINA 53也增加了LTR处H3 K36 me 3的局部水平。定位下游效应子的努力揭示了H3 K36 me 3与另一个表观遗传标记H4 K16 ac具有交叉作用，该标记由识别甲基化H3 K36和乙酰化H4 K16的KAT 8介导。去除MINA 53介导的潜伏机制可能有利于逆转整合后潜伏的HIV-1前病毒，以清除储库细胞。我们进一步证明了pan jumonji组蛋白去甲基化酶抑制剂JIB-04抑制MINA 53介导的H3 K36 me 3去甲基化，并且JIB-04与其他潜伏逆转剂（LRA）协同作用以重新激活潜伏的HIV-1。
Although combination antiretroviral therapy is potent to block active replication of HIV-1 in AIDS patients, HIV-1 persists as transcriptionally inactive proviruses in infected cells. These HIV-1 latent reservoirs remain a major obstacle for clearance of HIV-1. Investigation of host factors regulating HIV-1 latency is critical for developing novel antiretroviral reagents to eliminate HIV-1 latent reservoirs. From our recently accomplished CRISPR/Cas9 sgRNA screens, we identified that the histone demethylase, MINA53, is potentially a novel HIV-1 latency-promoting gene (LPG). We next validated MINA53's function in maintenance of HIV-1 latency by depleting MINA53 using the alternative RNAi approach. We further identified that in vitro MINA53 preferentially demethylates the histone substrate, H3K36me3 and that in cells MINA53 depletion by RNAi also increases the local level of H3K36me3 at LTR. The effort to map the downstream effectors unraveled that H3K36me3 has the cross-talk with another epigenetic mark H4K16ac, mediated by KAT8 that recognizes the methylated H3K36 and acetylated H4K16. Removing the MINA53-mediated latency mechanisms could benefit the reversal of post-integrated latent HIV-1 proviruses for purging of reservoir cells. We further demonstrated that a pan jumonji histone demethylase inhibitor, JIB-04, inhibits MINA53-mediated demethylation of H3K36me3, and JIB-04 synergizes with other latency-reversing agents (LRAs) to reactivate latent HIV-1.