Virus-Mosquito mRNA Stability

描述（由申请方提供）：蚊子是多种病毒病原体的载体，其中大量是RNA病毒。病毒控制的策略可以针对载体本身或病毒与载体之间的相互作用。该应用基于这样的假设，即RNA病毒已经进化出逃避蚊子细胞的mRNA周转机制的特定机制，因此mRNA衰变因子可能代表治疗的新靶点。我们目前对昆虫mRNA衰变的了解是最少的，因此本提案的主要目标必须是阐明伊蚊中mRNA周转所涉及的因素和途径，并表征其调节。使用体外的方法，我们已经成功地适应蚊子细胞提取物沿着在体内测定，目的我的目标是表征蚊子的mRNA去腺苷化，脱帽和腐烂的过程。在目的II中，我们将利用这些知识来深入了解我们观察到的由披膜病毒3'非翻译区介导的调节mRNA衰变的机制。该申请的最终目的将解决负责非多聚腺苷酸化病毒RNA如何在进入蚊子细胞后避免降解的潜在机制。总之，这些信息将提供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.