宿主因子PEX19与流感病毒M2蛋白互作影响病毒复制的分子机制研究

Influenza is an important zoonosis. The widely used clinical anti-influenza drugs are designed against influenza virus NA or M2 protein,which tends to drive the emergency of drug-resistant strains easily. Thus, it is of great significance to study the key host factors required for virus replication for discovery of new drug targets. M2 protein has many important functions in the cycle of influenza virus replication. Currently, very few host factors have been identified to interact with M2 protein. In previous work of this project, we have identified one host factor peroxisomal biogenesis factor 19 (PEX19) to interact with M2 protein by using the yeast two hybrid system (Y2H). Our results showed that PEX19 interact with M2 protein can affect the replication of influenza virus. On the basis of these results, we will further study the key residues for the interaction between the two proteins and the molecular mechanisms by which PEX19 regulate the replication of influenza virus. The implementation of the project will elucidate the molecular mechanism by which the PEX19 protein affects virus replication, deepen the understanding of influenza virus replication cycle, and providing new ideas for the development of new anti-influenza virus drugs.

流感是一种重要的人兽共患病。目前临床上广泛使用的抗流感药物是针对流感病毒NA蛋白或M2蛋白所设计，易于产生耐药毒株。深入研究流感病毒感染宿主细胞后复制过程中所依赖的宿主因子并阐明其调控病毒复制的分子机制，对于发现新的药物靶标具有重要意义。M2蛋白在流感病毒复制周期中具有多种重要功能，但目前已知与其互作的宿主因子很少。本课题在前期工作中，利用酵母双杂交系统筛选到一个同M2蛋白互作的宿主因子Peroxisomal biogenesis factor 19(PEX19)，我们已有研究结果表明M2蛋白同PEX19蛋白之间存在确切的相互作用且可以影响流感病毒复制，为进一步研究二者之间互作的关键区域及PEX19蛋白调控流感病毒复制的分子机制奠定了基础。通过本项目的实施将阐明PEX19蛋白同M2蛋白互作影响病毒复制的分子机制，深化对流感病毒复制周期的认识，为研制新型抗流感病毒药物提供一定新思路。

流感病毒M2蛋白在病毒复制过程中发挥多种重要功能，同时其功能的发挥离不开大量宿主因子的参与。在本项目研究过程中，我们利用酵母双杂交系统筛选到了同M2蛋白互作的宿主因子过氧化物酶体生物合成因子19 (PEX19)，我们的系统研究表明流感病毒感染宿主细胞过程M2蛋白同PEX19蛋白之间存在相互作用。通过构建M2蛋白及PEX19蛋白一系列截短表达突变体并进行Co-IP实验，明确了M2蛋白与PEX19蛋白互作的关键区域。利用构建的PEX19过表达细胞系、PEX19敲除细胞系及对照细胞系分别感染H1N1、H5N1及H7N9三种不同亚型流感病毒，我们发现PEX19蛋白对流感病毒复制发挥负调控作用，而且该调控作用对于不同亚型流感病毒均具有普遍性，初步探索了其相互作用的影响机制。通过本项目的实施，进一步深化我们对于宿主因子通过与M2蛋白互作调控流感病毒复制周期的认识。