NF-κB-Interacting Long Noncoding RNA Regulates HIV-1 Replication and Latency by Repressing NF-κB Signaling
NF-κB-Interacting Long Noncoding RNA Regulates HIV-1 Replication and Latency by Repressing NF-κB Signaling
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NF-kappa B 相互作用长非编码 RNA 通过抑制 NF-kappa B 信号传导调节 HIV-1 复制和潜伏期
DOI：
10.1128/jvi.01057-20
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发表时间：
2020-09-01
影响因子：
5.4
通讯作者：
Zhang, Wenyan
中科院分区：
文献类型：
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作者：
Wang, Hong;Liu, Yue;Zhang, Wenyan
NF-kappa B相互作用的长链非编码RNA （NKILA）最近被发现是NF-kappa B信号传导的负调控因子，在各种癌症的发展中发挥重要作用。众所周知，NF-kappa b介导的人类免疫缺陷病毒1型（HIV-1）长末端重复（LTR）驱动基因表达的激活是HIV-1转录和潜伏期再激活所必需的。然而，NKILA是否在HIV-1复制和潜伏期中发挥重要作用尚不清楚。在这里，通过异位表达和沉默实验，我们证明NKILA通过抑制HIV-1 LTR启动子活性，以nf - κ b依赖的方式有效抑制HIV-1复制。此外，NKILA对具有不同共受体趋向性的HIV-1克隆的复制以及各种HIV-1临床亚型的LTR活性具有广谱抑制作用。染色质免疫沉淀（ChIP）分析显示，NKILA的表达消除了p65在HIV-1 LTR中复制的KB结合位点的募集，破坏NF-kappa B抑制的NKILA突变体也失去了抑制HIV-1复制的能力。值得注意的是，HIV-1感染或再激活显著下调了T细胞中NKILA的表达，以促进病毒复制。NKILA的下调主要是由于HIV-1感染降低了NKILA启动子上组蛋白K27的乙酰化，从而阻断了NKILA的表达。在体外建立的azidothymidine （AZT）停药临床模型中，敲低NKILA可促进潜伏性HIV-1在肉豆酸酯磷（phorbol myristate acetate， PMA）刺激下的再激活，而异位NKILA则抑制其再激活。这些发现提高了我们对NKILA通过NF-kappa B信号传导对HIV-1复制和潜伏期的功能性抑制的理解。nf - κ B通路在HIV-1复制和HIV-1潜伏期的再激活中起关键作用。抑制nf - κ B活化的调节因子可能是一种有希望的治疗HIV-1的策略。最近，NF-kappa B相互作用的长链非编码RNA （NKILA）被发现通过抑制IKK诱导的I -kappa B磷酸化和NF-kappa B通路激活来抑制不同人类癌症的发展，而NKILA与HIV-1复制的关系尚不清楚。在这里，我们的研究结果表明NKILA通过抑制HIV-1长末端重复（LTR）驱动的转录起始来抑制HIV-1的复制和再激活。此外，NKILA抑制了具有不同共受体趋向性的HIV-1克隆的复制。该项目可能为开发新型抗hiv药物提供一个靶点。
NF-kappa B-interacting long noncoding RNA (NKILA) was recently identified as a negative regulator of NF-kappa B signaling and plays an important role in the development of various cancers. It is well known that NF-kappa B-mediated activation of human immunodeficiency virus type 1 (HIV-1) long terminal repeat (LTR)-driven gene expression is required for HIV-1 transcription and reactivation of latency. However, whether NKILA plays essential roles in HIV-1 replication and latency is unclear. Here, by ectopic expression and silencing experiments, we demonstrate that NKILA potently inhibits HIV-1 replication in an NF-kappa B-dependent manner by suppressing HIV-1 LTR promoter activity. Moreover, NKILA showed broad-spectrum inhibition on the replication of HIV-1 clones with different coreceptor tropisms as well as on LTR activity of various HIV-1 clinical subtypes. Chromatin immunoprecipitation (ChIP) assays revealed that NKILA expression abolishes the recruitment of p65 to the duplicated KB binding sites in the HIV-1 LTR. NKILA mutants disrupting NF-kappa B inhibition also lost the ability to inhibit HIV-1 replication. Notably, HIV-1 infection or reactivation significantly downregulated NKILA expression in T cells in order to facilitate viral replication. Downregulated NKILA was mainly due to reduced acetylation of histone K27 on the promoter of NKILA by HIV-1 infection, which blocks NKILA expression. Knockdown of NKILA promoted the reactivation of latent HIV-1 upon phorbol myristate acetate (PMA) stimulation, while ectopic NKILA suppressed the reactivation in a well-established clinical model of withdrawal of azidothymidine (AZT) in vitro. These findings improve our understanding of the functional suppression of HIV-1 replication and latency by NKILA through NF-kappa B signaling.IMPORTANCE The NF-kappa B pathway plays key roles in HIV-1 replication and reactivation of HIV-1 latency. A regulator inhibiting NF-kappa B activation may be a promising therapeutic strategy against HIV-1. Recently, NF-kappa B-interacting long noncoding RNA (NKILA) was identified to suppress the development of different human cancers by inhibiting I kappa B kinase (IKK)-induced I kappa B phosphorylation and NF-kappa B pathway activation, whereas the relationship between NKILA and HIV-1 replication is still unknown. Here, our results show that NKILA inhibits HIV-1 replication and reactivation by suppressing HIV-1 long terminal repeat (LTR)-driven transcription initiation. Moreover, NKILA inhibited the replication of HIV-1 clones with different coreceptor tropisms. This project may reveal a target for the development of novel anti-HIV drugs.