鉴定参与HEV复制的宿主蛋白及其机制研究

戊型肝炎病毒（HEV）的感染能引起急性肝炎、慢性感染和孕妇的高死亡率。目前，市面上没有治疗HEV感染的特效药，科学家对参与HEV复制的宿主蛋白及病毒复制机制也知之甚少。基于全长的HEV复制酶不需要加工成独立的蛋白就有复制能力，本项目计划用HEV复制酶反式互补系统，通过细胞培养稳定同位素标记技术（SILAC）和质谱技术鉴定与其相互作用的宿主蛋白；同时，通过构建HA-Flag标签标记ORF1的病毒复制子，分析HEV复制复合体的组成，在加深对病毒复制机制理解的同时，也为开发靶向宿主蛋白的抗病毒药物提供靶点。前期研究发现，PRMT5和WDR77能够抑制HEV复制，通过探究包括PRMT5和WDR77在内的宿主因子调控HEV复制的机制，为理解病毒与宿主相互作用和治疗HEV感染提供依据。

Hepatitis E virus (HEV) infection can cause acute hepatitis, chronic infection and high mortality in pregnant women. Currently, there is no direct antiviral agent to treat HEV infection on the market, and little is known about the host proteins involved in and the mechanism of HEV replication. The full-length HEV replicase does not need to be processed into different independent proteins to replicate viral RNA. In this project, we plan to utilize the HEV replicase trans-complementation system to identify the host proteins that interact with ORF1 by stable isotope labeling with amino acids in cell culture (SILAC) and mass spectrometry. At the same time, by constructing the viral replicon of ORF1 tagged with HA-Flag tag, the HEV replication complex will be analyzed. This not only deepens our understanding of viral replication mechanism, but also provides targets for developing antivirals. Previously, we have found that PRMT5 and WDR77 can inhibit HEV replication. Exploring the mechanism by which host factors including PRMT5 and WDR77 regulate HEV replication will provide a basis for understanding virus-host interactions and treating HEV infection.