P小体在维持胞内RNA水平的动态平衡及翻译过程中发挥着重要作用，病毒RNA的转录、翻译、定位及降解过程能够与P小体相互作用，影响病毒复制。P小体标志蛋白GW182在病毒感染复制过程中的作用知之甚少。我们的预实验结果发现，柯萨奇病毒B3（CVB3）感染细胞后，病毒RNA能够与GW182在胞质中共定位，同时在细胞分泌的外泌体中也共定位；敲除GW182可抑制CVB3病毒复制。据此我们提出假说：P小体标志蛋白GW182能够参与CVB3病毒的自身复制，同时影响病毒RNA通过外泌体途径释放。围绕这一假说，我们将运用免疫共沉淀、RNA pull down、CLIP-Seq及基因敲除动物模型等技术手段，阐明1）GW182在CVB3病毒复制中的作用及分子机制；2）GW182在CVB3病毒RNA外泌体释放中的作用及病理意义。本课题将为病毒如何利用P小体标志蛋白GW182来影响自身复制和胞外传播致病提供新证据。

P-bodies play an important role in maintaining the dynamic balance of intracellular mRNA levels and the process of intracellular translation. Viral RNA can interact with P-bodies for transcription, translation, localization and degradation, as well as viral replication. Little is known about the role of the P-bodies marker GW182 during virus replication. Our preliminary data showed that the viral RNA of CVB3 could co-localize with GW182 protein in CVB3-infected cells，similar result was also found in the exosomes that from virus-infected cells. Knockdown of GW182 protein could inhibit CVB3 RNA replication. Therefore, we hypothesize that the GW182 is involved in CVB3 viral replication, as well as can affect the viral RNA release through exosomes pathway. We will test this hypothesis by using Co-Immunoprecipitation, RNA-pull down, CLIP-Seq and Transgenic and Knockout Animal models. We will investigate that (1)The role of P-bodies marker GW182 protein during CVB3 replication, as well as the molecular mechanism.(2) The function of GW182 in viral RNA release through exosomes process. This program will provide innovative evidence for how the virus uses the P-bodies marker GW182 to affect its own replication and the pathogenesis of extracellular transmission.