甲型流感病毒NS1蛋白与宿主蛋白eIF3f相互作用促进病毒复制的研究

Interaction between influenza A virus NS1 protein and host protein eIF3f promotes viral replication

甲型流感病毒(IAV)非结构蛋白1(NS1)是一种重要的毒力因子,其通过与多种宿主细胞因子相互作用抑制宿主细胞天然免疫反应和调控流感病毒复制。为筛选与NS1相互作用的宿主蛋白,本研究利用H1N1亚型流感病毒NS1蛋白作为“诱饵”蛋白,经过Flag pull-down联合质谱分析筛选得到4个与NS1可能相互作用的宿主蛋白eIF3f、ENO1、CCAR2和PSB2。对筛选得到的宿主蛋白评分分析后,针对宿主蛋白eIF3f进行双分子荧光互补分析(BiFC)试验验证NS1蛋白与eIF3f蛋白的相互作用,结果显示eIF3f蛋白与NS1蛋白存在相互作用,并且在HEK293T细胞中瞬时过表达的eIF3f促进了H1N1亚型流感病毒的复制。本研究为进一步探究eIF3f蛋白在流感病毒复制中发挥的生物学功能奠定了基础。

The nonstructural protein 1(NS1)of influenza A virus(IAV)is a central virulence factor,which can inhibit host innate immune response and regulate virus replication by interacting with multiple host cell factors.In the present study,we chose the NS1 protein of 2009 H1N1 influenza virus to construct a bait protein for screening host cellular proteins.By using Flag pull-down and Mass Spectrometric analysis,several proteins including eIF3f(Eukaryotic translation initiation factor 3 subunit F),ENO1(MBP-1 of Alpha-enolase),CCAR2(Cell cycle and apoptosis regulator protein 2)and PSB2(Proteasome subunit beta type-2)were potentially interacted with NS1 protein.After analyzing the score of screened proteins,thehost protein eIF3f was selected for further study,and the result showed that specific interaction between NS1 and eIF3f was identified by Bimolecular Fluorescence Complementation assay.Overexpression of eIF3f promoted H1N1 subtype virus replication in HEK293T cells.These data thus provide a basis to further study the biological function of eIF3f protein in the replication process of influenza A virus.