A novel antiviral approach using the cellular RNA decay machinery

细胞RNA衰变机制通常会对细胞进行监管，并有效地去除不需要的RNA。我们 最近发现，包括VEE在内的甲型病毒具有特定的RNA稳定元件 它们的3‘端非编码区招募一种细胞因子，阻止病毒转录本的去烯化/衰变，从而 促进高效和多产的感染。根据这些数据，我们假设，即使不是全部，也有很多 RNA病毒编码有帽子结构和多聚腺苷化的转录本，已经进化出选择性地 抑制细胞信使核糖核酸的衰退机制是有效表达病毒基因的先决条件。目标是 这项提议的目的是通过评估我们是否可以扩大我们的观察范围来检验这一假设 对附加甲型病毒的细胞RNA去烯化/衰退机制的病毒抑制(WEE和 EEE)以及负义RNA病毒(埃博拉、马尔堡和尼帕)。除了扩大我们的 分子宿主-病毒相互作用的知识，这些研究还将测试这些试剂是否使用 与甲型病毒类似的调节病毒RNA稳定性的策略。如果这些RNA病毒使用单个(AS 初步数据表明，甲型病毒的情况)或有限数量的策略来抑制细胞 MRNA衰变机制，这是抗病毒治疗的一个新的和有吸引力的靶点。为此， 目标2的目标是优化我们已建立的用于测量病毒mRNA稳定性的体外检测方法，以允许 化合物文库的快速有效筛选。这项提案的最终目标是执行 用于识别和验证可克服病毒抑制的候选先导化合物的化学文库筛选 细胞核糖核酸衰变机制。这些化合物将是更具吸引力的候选化合物 作为抗病毒治疗药物的开发，很可能对各种 具有生物防御意义的病毒。该研究项目符合RMRCE综合研究重点 关于病毒治疗，并将直接与RPS 3.1、3.2、3.5、3.7和3.8互动，并利用资源 这个项目的目标应该为RMRCE增加重要的专业知识，并与之很好地协同 病毒治疗焦点小组，特别是由于提出了创新的基本病毒学问题和 开发可以针对多个RNA病毒家族的抗病毒药物的新方法 组。

The cellular RNA decay machinery routinely polices the cell and effectively removes unwanted RNAs. We have recently discovered that alphaviruses, including VEE, possess specific RNA stabilizing elements in their 3' UTR that recruit a cellular factor and block the deadenylation/decay of viral transcripts, thereby promoting an efficient and productive infection. Based on these data, we hypothesize that many if not all RNA viruses that encode capped and polyadenylated transcripts have evolved mechanisms to selectively suppress the cellular mRNA decay machinery as a prerequisite for efficient virus gene expression. The goal of Aim I of this proposal is to test this hypothesis by assessing whether we can extend our observations on viral suppression of the cellular RNA deadenylation/decay machinery to additional alphaviruses (WEE and EEE) as well as negative sense RNA viruses (Ebola, Marburg and Nipah). In addition to expanding our knowledge of molecular host-virus interactions, these studies will also test whether these agents employ a similar strategy as the alphaviruses for mediating viral RNA stability. If these RNA viruses use a single (as preliminary data indicate is the case for alphaviruses) or limited number of strategies to suppress the cellular mRNA decay machinery, this represents a novel and attractive target for antiviral therapeutics. To this end, the goal of Aim 2 is to optimize our established in vitro assays for measuring viral mRNA stability to allow for the rapid and effective screening of chemical compound libraries. The final aim of this proposal is to perform a chemical library screen to identify and validate candidate lead compounds that overcome viral suppression of the cellular RNA decay machinery. These compounds would represent attractive candidates for further development as antiviral therapeutics that may very well have a broad spectrum activity against a variety of viruses of biodefense significance. This research project fits within the RMRCE Integrated Research Focus on Viral Therapeutics, and will interact directly with RPs 3.1, 3.2, 3.5, 3.7 and 3.8, and utilize the resources of Core E. The goals of this project should add significant expertise to and synergize well with the RMRCE Viral Therapeutics Focus Group, particularly due to the innovative basic virology questions being asked and the novel approach to develop antivirals that could target multiple families of RNA viruses of interest to the Group.