Xu Qiang, Huang Lihong, Xing Jinchao, Zhang Jiahao, Li Huanan, Liu Lele, Hu Chen, Liao Ming, Yue Jianbo, Qi Wenbao
College of Veterinary Medicine, South China Agricultural University, Guangzhou, 510642, China; Key Laboratory of Zoonosis, Ministry of Agriculture and Rural Affairs, Guangzhou, 510642, China; National and Regional Joint Engineering Laboratory for Medicament of Zoonosis Prevention and Control, Guangzhou, 510642, China; Key Laboratory of Zoonoses Prevention and Control of Guangdong Province, Guangzhou, 510642, China.
Department of Biomedical Sciences, City University of Hong Kong, 999077, Hong Kong, China.
Vet Microbiol. 2021 Apr;255:109025. doi: 10.1016/j.vetmic.2021.109025. Epub 2021 Mar 8.
Japanese encephalitis virus is absolutely dependent on their host cells and has evolved various strategies to manipulate the cellular secretory pathways for viral replication. However, how cellular secretory pathways are hijacked, and the origin of the viral vesicles remains elusive during JEV replication. Here we show how JEV manipulates multiple components of the cellular secretory pathway, including autophagic machinery, to generate a superior environment for genome replication. We utilized double-strand RNA antibodies to label JEV RNA complex seeking the viral replication compartments and found that JEV genome replication takes place in lysosomes (LAMP1), not in autophagosomes (LC3). Subsequently, in situ hybridization results showed that viral RNAs (vRNAs) of JEV strongly colocalized with LAMP1. What surprised us was that JEV vRNAs markedly colocalized with LC3, indicating that autophagy plays an active role in JEV replication. Interestingly, we found that JEV utilized autophagic components for intracellular growth in an autophagy-dependent manner and the fusion of autophagosome-lysosome plays a positive role in JEV post-RNA replication processes. Collectively, our findings demonstrate that JEV can manipulate cellular secretory pathway to form genome replication organelles and exploit autophagy components for intracellular growth, providing new insights into the life cycle of JEV and uncovering an attractive target for antiviral drugs.
日本脑炎病毒完全依赖其宿主细胞，并已进化出多种策略来操纵细胞分泌途径以进行病毒复制。然而，在日本脑炎病毒复制过程中，细胞分泌途径是如何被劫持的，以及病毒囊泡的起源仍然不清楚。在这里，我们展示了日本脑炎病毒如何操纵细胞分泌途径的多个组成部分，包括自噬机制，以创造一个有利于基因组复制的环境。我们利用双链RNA抗体标记日本脑炎病毒RNA复合物以寻找病毒复制区室，发现日本脑炎病毒基因组复制发生在溶酶体（LAMP1）中，而不是自噬体（LC3）中。随后，原位杂交结果显示，日本脑炎病毒的病毒RNA（vRNA）与LAMP1强烈共定位。令我们惊讶的是，日本脑炎病毒vRNA与LC3明显共定位，表明自噬在日本脑炎病毒复制中发挥积极作用。有趣的是，我们发现日本脑炎病毒以自噬依赖的方式利用自噬成分进行细胞内生长，并且自噬体-溶酶体的融合在日本脑炎病毒RNA复制后过程中发挥积极作用。总的来说，我们的研究结果表明，日本脑炎病毒可以操纵细胞分泌途径以形成基因组复制细胞器，并利用自噬成分进行细胞内生长，为日本脑炎病毒的生命周期提供了新的见解，并揭示了一个有吸引力的抗病毒药物靶点。
