Abstract:
The replication of Ebola virus (EBOV) is dependent upon actin functionality, especially at cell entry through macropinocytosis and at release of virus from cells. Previously, major actin-regulatory factors involved in actin nucleation, such as Rac1 and Arp2/3, were shown important in both steps. However, downstream of nucleation, many other cell factors are needed to control actin dynamics. How these regulate EBOV infection remains largely unclear. Here, we identified the actin-regulating protein, CAPG, as important for EBOV replication. Notably, knockdown of CAPG specifically inhibited viral infectivity and yield of infectious particles. Cell-based mechanistic analysis revealed a requirement of CAPG for virus production from infected cells. Proximity ligation and split-green fluorescent protein reconstitution assays revealed strong association of CAPG with VP40 that was mediated through the S1 domain of CAPG. Overall, CAPG is a novel host factor regulating EBOV infection through connecting actin filament stabilization to viral egress from cells.
摘要:
埃博拉病毒(EBOV)的复制依赖于肌动蛋白的功能,特别是在通过巨细胞胞吞作用进入细胞和从细胞释放病毒时。以前,参与肌动蛋白成核的主要肌动蛋白调节因子,如Rac1和Arp2/3,在这两个步骤中都显示出重要的作用。然而,成核的下游,许多其他细胞因子需要控制肌动蛋白动力学。这些如何调节EBOV感染仍不清楚。这里,我们确定了肌动蛋白调节蛋白,CAPG,对EBOV复制很重要。值得注意的是,CAPG的敲低特异性地抑制病毒感染性和感染性颗粒的产量。基于细胞的机理分析揭示了从感染细胞产生病毒需要CAPG。邻近连接和分裂绿色荧光蛋白重建测定揭示了CAPG与VP40的强关联,其通过CAPG的S1结构域介导。总的来说,CAPG是一种新型宿主因子,通过将肌动蛋白丝稳定与病毒从细胞中排出来调节EBOV感染。