1Vaccine and Infectious Disease Research Centre, Translational Health Science and Technology Institute, NCR Biotech Science Cluster, Faridabad, Haryana, India.
2Symbiosis School of Biological Sciences, Symbiosis International (Deemed University), Pune, Maharashtra, India.
J Gen Virol. 2019 Feb;100(2):176-186. doi: 10.1099/jgv.0.001182. Epub 2018 Nov 29.
Japanese encephalitis virus (JEV), a mosquito-borne flavivirus, is one of the leading global causes of virus-induced encephalitis. The infectious life-cycle of viruses is heavily dependent on the host membrane trafficking network. Here, we have performed a RNA-interference-based screen using a siRNA panel targeting 136 membrane trafficking proteins to identify the key regulators of JEV infection in HeLa cells. We identified 35 proteins whose siRNA depletion restricts JEV replication by over twofold. We observe that JEV infection in HeLa cells is largely dependent on components of the clathrin-mediated endocytic (CME) pathway. Proteins involved in actin-filament-based processes, specifically CDC42 and members of the ARP2/3 complex are crucial for establishment of infection. Pharmacological pertubations of actin polymerization, a small molecule inhibitor of actin nucleation by the ARP2/3 complex - CK-548 - and the inhibitor of neural Wiskott-Aldrich syndrome proteins- Wiskostatin- inhibited JEV replication, highlighting the important role of the dynamic actin network. Other proteins involved in cargo-recognition for CME and endomembrane system organization were also validated as essential host factors for virus replication.
日本脑炎病毒（JEV）是一种通过蚊子传播的黄病毒，是导致病毒性脑炎的主要全球病因之一。病毒的感染生命周期严重依赖于宿主的膜转运网络。在这里，我们使用靶向 136 种膜转运蛋白的 siRNA 面板进行了基于 RNA 干扰的筛选，以鉴定 HeLa 细胞中 JEV 感染的关键调节剂。我们鉴定出 35 种蛋白质，其 siRNA 耗尽会使 JEV 复制超过两倍。我们观察到 JEV 在 HeLa 细胞中的感染在很大程度上依赖于网格蛋白介导的内吞作用（CME）途径的成分。肌动蛋白丝基于过程的蛋白质，特别是 CDC42 和 ARP2/3 复合物的成员，对于建立感染至关重要。肌动蛋白聚合的药理学扰动、ARP2/3 复合物的肌动蛋白成核的小分子抑制剂 CK-548 和神经 Wiskott-Aldrich 综合征蛋白抑制剂 Wiskostatin 抑制了 JEV 的复制，突出了动态肌动蛋白网络的重要作用。其他参与 CME 和内膜系统组织货物识别的蛋白质也被验证为病毒复制的必需宿主因子。
