Functional Influenza NS1-Cellular Protein Interactions

描述：（改编自《研究者》摘要）：本研究旨在调查甲型流感和乙型流感NS1蛋白在病毒感染过程中与特定细胞蛋白之间的功能相互作用。ns1蛋白是一种非结构蛋白，对a型病毒来说，它调控转录后事件。nslb蛋白的功能尚不清楚。反向遗传学的最新进展使甲型和乙型流感病毒完全由克隆的cdna产生。这是产生突变型流感病毒和研究流感感染过程中发生的蛋白质相互作用的有力工具。该研究利用该技术以及GST下拉、亲和层析、突变和缺失分析来鉴定与ns1蛋白相互作用的特定细胞蛋白。这些蛋白质相互作用将为开发更有效的流感抗病毒疗法提供见解。具体目的是：1)确定特定细胞蛋白与甲型流感病毒ns1a蛋白之间的功能相互作用，重点确定这种相互作用与SRp54的功能。将鉴定蛋白质结合位点，并分析这种特异性蛋白质相互作用的功能。2)确定特定细胞蛋白与乙型流感病毒NS lB蛋白之间的功能相互作用。将鉴定蛋白质结合位点，并分析这些特定蛋白质相互作用的功能。重点将放在nslb和ISG15之间的相互作用。

DESCRIPTION: (Adapted from the Investigators abstract): The proposed research investigates functional interactions between the influenza A and B NS1 proteins with specific cellular proteins during viral infection. The NS 1 protein is a non-structural protein that for type A viruses regulates events post-transcription. The function(s) of the NS lB protein remains unclear. Recent advances in reverse genetics allow influenza A and B viruses to be generated entirely from cloned cDNAs. This is a powerful tool for the generation of mutant influenza viruses and the investigation of protein interactions that occur during the course of influenza infection. The proposed research utilizes this technique as well as GST pull-downs, affinity chromatography, and mutation and deletion analysis to identify specific cellular proteins that interact with the NS 1 protein. These protein interactions will provide insight that could allow for the development of more effective influenza antiviral therapies. Specific aims are: 1) Identify functional interactions between specific cellular proteins and the NS 1A protein of influenza A virus, with emphasis on identifying the function of this interaction with SRp54. The protein-binding site will be identified and the function of this specific protein interaction analyzed. 2) Identify functional interactions between specific cellular proteins and the NS lB protein of influenza B virus. The protein-binding sites will be identified and the function of these specific protein interactions analyzed. Emphasis will be on the interaction between NS lB and ISG15.