Virus-Mosquito mRNA Stability

描述（申请人提供）：蚊子是多种病毒性病原体的载体，其中大量是RNA病毒。病毒控制策略可以针对载体本身或病毒与载体之间的相互作用。这一应用是基于这样的假设，即RNA病毒已经进化出特定的机制来逃避蚊子细胞的mRNA转换机制，因此mRNA衰变因子可能代表一种新的治疗靶点。我们目前对昆虫mRNA衰变的了解很少，因此本提案的主要目标必须是阐明伊蚊mRNA转换的因素和途径，并表征其调控。利用我们已经成功适应于蚊子细胞提取物的体外方法以及体内试验，Aim I的目标是表征蚊子mRNA死基化，脱冠和衰变的过程。在Aim II中，我们将利用这些知识来深入了解我们观察到的托加病毒3'非翻译区介导的受调节的mRNA衰变机制。该应用程序的最终目的是解决非聚腺苷化病毒rna在进入蚊子细胞时如何避免降解的潜在机制。总之，这些信息将为昆虫分子生物学和病毒-宿主相互作用提供基本见解，为虫媒病毒控制的新途径提供基础。

DESCRIPTION (provided by applicant): Mosquitoes are vectors of multiple viral pathogens, a large number of which are RNA viruses. Strategies for viral control may target the vector itself or the interaction between the virus and the vector. This application is based upon the hypothesis that RNA viruses have evolved specific mechanisms for evading the mRNA turnover machineries of the mosquito cell and therefore mRNA decay factors may represent a novel target for therapeutics. Our current knowledge of mRNA decay in insects is minimal thus the primary goals of this proposal must be to elucidate the factors and pathways involved in mRNA turnover in the Aedes mosquito and characterize their regulation. Using an in vitro approach that we have successfully adapted to mosquito cell extracts along with in vivo assays, the goal of Aim I is to characterize the processes of mRNA deadenylation, decapping and decay in mosquitoes. In Aim II we will use this knowledge to gain insights into mechanisms of regulated mRNA decay mediated by togavirus 3' untranslated regions that we have observed. The final aim of the application will address the underlying mechanisms responsible for how non- polyadenylated viral RNAs avoid degradation upon entry into the mosquito cell. In summary, this information will provide fundamental insights into insect molecular biology and virus-host interactions that will provide the groundwork for novel avenues of arbovirus control.