人类免疫缺陷病毒(HIV)编码的RNA结合蛋白Rev对宿主转录组的调节作用

HIV is a major global public health threat. So far there is no cure for its infection. And even the most effective treatment, HAART, is facing major challenges from both severe side effects and the emergence of drug-resistant viral strains. To facilitate the development of alternative therapies, continuous efforts in understanding the mechanistic details of viral life events would be essential. Understanding the complex interactions between HIV and host components can offer tremendous insights into both HIV replication and virus-host interplay. Furthermore, due to the rapid mutation of the virus itself, host factors critical to viral replication become promising antiviral targets. Using a series of systems-wide methods like PAR-CLIP and RNA-Seq, we will target the HIV RNA binding protein, Rev, and set out to reveal the landscape of its complex interactions with the host transcriptome. Furthermore, using a combination of biochemical, molecular and structural biology methods, we will further demonstrate how Rev regulates two specific types of host RNAs identified in our preliminary studies. First, analogous to its canonical function, we will investigate how Rev interacts with host intron-containing mRNAs to facilitate their nuclear-export and alter their functions in host immune response. Second, we will characterize the interactions between Rev and human RPPH1, which is the RNA component of RNase P. We will also further demonstrate the connection between this interaction and the change of cellular tRNA composition. This study will greatly enrich our knowledge for different layers of virus-host interactions and bring new insights to support the development of new therapies.

HIV对世界公共卫生产生严重威胁，其感染尚无根治方法，而最佳控制手段HAART也面临两大挑战：副作用及抗药型病毒株的涌现。为推动新型治疗的研发，我们迫切需要对病毒复制过程做更深入的了解。其中，对病毒与宿主分子复杂结合网络的测定能加强我们对病毒复制机理的了解。而HIV的高突变率使得对其复制有决定作用的宿主分子成为制药的重要靶向。本项目将针对HIV的RNA结合蛋白Rev，运用PAR-CLIP、RNA-Seq等方法，揭示其同人类RNA的结合网络及其对转录组成分的系统调控。此外，还将深入分析Rev与两类宿主RNA的代表性相互作用。一方面，将揭示Rev介导包涵内含子的宿主mRNA出核运输的机理及其对细胞免疫反应的影响；另一方面，将揭示Rev同非编码RPPH1 RNA结合并进一步调节细胞内的tRNA谱的机制。本项目从新的角度揭示病毒与宿主的关联，将为新药的研发奠定坚实基础，具有重大科研和应用意义。

HIV对世界公共卫生产生严重威胁，其感染尚无根治方法，而最佳控制手段HAART也面临两大挑战：副作用及抗药型病毒株的涌现。为推动新型治疗的研发，我们迫切需要对病毒复制过程做更深入的了解。本项目研究目标是HIV编码的RNA结合蛋白Rev和Gag对宿主及病毒RNA的调控。其中，对病毒与宿主分子复杂结合网络的测定能加强我们对病毒复制机理的了解。而HIV的高突变率使得对其复制有决定作用的宿主分子成为制药的重要靶向。针对病毒蛋白Rev与宿主转录组的相互作用，我们首先运用PAR-CLIP、RNA-Seq等方法，揭示了Rev同人类细胞转录组的结合网络及其对转录组成分的系统调控。进而，我们深入研究并揭示了Rev介导的包涵内含子的宿主mRNA出核运输的机理及其对细胞免疫反应的影响。针对病毒蛋白Gag对病毒RNA的调控，我们运用EMSA、SHAPE等生物化学方法揭示了Gag与病毒RRE RNA结合的位点以及结合时RNA的结构变化过程。进而，我们揭示在病毒RNA翻译过程中，RRE结合的Rev蛋白可以被有效剔除，这为Gag-RRE的结合创造了条件。本项目从新的角度揭示病毒与宿主的关联，将为新药的研发奠定坚实基础，具有重大科研和应用意义。