Virus-Mosquito mRNA Stability

描述(申请人提供)：蚊子是多种病毒病原体的媒介，其中大量是RNA病毒。病毒控制策略可能针对媒介本身或病毒与媒介之间的相互作用。这一应用是基于这样一种假设，即RNA病毒已经进化出特定的机制来逃避蚊子细胞的mRNA周转机制，因此mRNA衰变因子可能代表着治疗的新靶点。我们目前对昆虫中的mRNA衰变的了解很少，因此这项提议的主要目标必须是阐明参与伊蚊mRNA更替的因素和途径，并描述它们的调节。使用我们已经成功适应蚊子细胞提取物的体外方法以及体内测试，目标I的目标是表征蚊子中mRNA去烯化、去帽和腐烂的过程。在AIM II中，我们将利用这些知识来深入了解我们观察到的togaVirus 3‘非翻译区介导的受调控的mRNA衰退的机制。该应用程序的最终目标将解决导致非多腺化病毒RNA如何在进入蚊子细胞时避免降解的潜在机制。总之，这些信息将提供对昆虫分子生物学和病毒-宿主相互作用的基本见解，这将为虫媒病毒控制的新途径奠定基础。

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.