很多正链RNA病毒重塑细胞内膜系统形成特殊的复制结构。然而负链RNA病毒是否也利用细胞内膜系统完成复制却知之甚少。申请人所在实验室以前的研究结果表明：人副流感病毒（HPIV3）的N-P相互作用调节病毒RNA的合成（Virology, 2018；Viruses, accepted）；包涵体融合形成成熟的包涵体是病毒RNA合成所必须的（Journal of Virology, 2017）；包涵体通过隐藏其合成的RNA逃逸宿主细胞的抗病毒反应（PLoS Pathogens, accepted）。在这些研究结果的基础上，我们期望揭示HPIV3的包涵体是否也利用细胞的内膜系统？包涵体发挥功能是否需要其他细胞因子的协助？本研究将通过免疫荧光、蛋白质与蛋白质相互作用、质谱分析、膜提取纯化和免疫电镜等技术，深入解析包涵体的组成、鉴定包涵体发挥功能所需的细胞因子，从而澄清包涵体调节病毒RNA合成的分子机制。

Many positive-stranded RNA viruses reorganized endo-membranes to form specialized replication organelles. However, it is largely unknown whether negative-stranded RNA viruses utilize intracellular membranes for their replication. Our previous studies showed that Human parainfluenza virus type 3 (HPIV3), a typical negative-strand RNA virus, replicates in inclusion bodies (IBs) (Virology, 2018; Viruses, accepted); Fusion of IBs is indispensable for replication of HPIV3; IBs can escape from antiviral activity of host cells by concealing viral RNA. Based on these results, we sought to explore whether IBs are the organelles derived from or utilize endo-membranes of host? In this study, we will identify and characterize the composition of IBs, and screen out the essential cell factors involving in viral replication via immunofluorescence, protein-protein interaction, mass-spectrographic, immune-electron microscopy and membrane composition extraction and purification. In summary, our study will further clarify how IBs accomplish the mission in HPIV3 replication.