Proteomics of HCV Replication Complex

描述(申请人提供)：丙型肝炎病毒(丙型肝炎病毒)慢性感染是导致重型肝炎的主要原因，重型肝炎通常发展为肝硬变和肝细胞癌。丙型肝炎病毒复制和致病的分子机制尚不清楚。与丙型肝炎病毒感染相关的肝病状态相关的宿主或病毒蛋白的身份尚不清楚。尽管我们最近发现了与丙型肝炎病毒3‘端非编码区相互作用的各种细胞因子(附录1)，但对丙型肝炎病毒复制复合体的组成成分知之甚少。这些成分有望在产生具有感染性的丙型肝炎病毒粒子过程中发挥关键作用。我们设计了一种从丙型肝炎病毒感染细胞中原位捕获复制复合体并通过蛋白质组学技术鉴定其成分的方法。我们将通过siRNA介导的基因敲除细胞中的丙型肝炎病毒表达来研究每个已识别的细胞成分对丙型肝炎病毒复制的影响。这将产生关于与复制复合体相关的细胞或病毒因子的身份的有价值的信息，并有助于识别用于预防病毒感染的新靶点。

DESCRIPTION (provided by applicant): Chronic infection by hepatitis C virus (HCV) is the leading cause of severe hepatitis, which often develops into liver cirrhosis and hepatocellular carcinoma. The molecular mechanisms underlying HCV replication and pathogenesis are poorly understood. The identity of host or viral proteins relevant to the state of liver disease associated with HCV infection is not known. Although we have recently identified various cellular factors that interact with HCV 3'NTR (Appendix #1), nothing is known about the components of the HCV replication complex. These components are expected to have crucial roles in the production of infectious HCV virions. We have devised a means of capturing the replication complex in situ from HCV-infected cells and identifying its components by proteomics technology. We will investigate the implication of each identified cellular component on HCV replication by siRNA-mediated genetic knockdown of its expression in the cells. This will yield valuable information about the identity of the cellular or viral factors associated with the replication complex and facilitate the identity of new targets for use in preventing the viral infection.