Characterization of host proteins that participate in the replication of hepatitis C virus (HCV)

丙型肝炎病毒（HCV）是人类肝脏疾病的主要病原体，目前尚无有效的疫苗接种程序，而且目前的治疗对大量患者无效。因此，阐明病毒复制过程的分子基础对于理解病毒发病机制和改善丙型肝炎的治疗非常重要。我们的建议旨在识别和功能特性的细胞蛋白质，其中，除了迄今已知的“宿主因子”，支持复制的HCV RNA基因组在细胞的细胞质中，并实现进一步了解病毒复制复合物（RC）的物理组织。最近的研究揭示了存在高分子量的胞质复合物，其含有HCV宿主因子如hVAP-A蛋白和所谓的NFAR蛋白以及各种未确定的蛋白。考虑到该复合物可能是病毒RC的中间体，将对其进行纯化，并通过质谱分析鉴定其蛋白质组分。将测试新识别的蛋白质在HCV RNA细胞内翻译和复制周期中的潜在作用，并通过遗传和生化手段分析蛋白质-蛋白质相互作用的分子细节。沿着这些路线，我们期望获得重要的信息组成的病毒复制机制，并扩大我们的知识的HCV的致病特征所造成的病毒招募的细胞成分。

Hepatitis C virus (HCV) is a major agent of liver disease in man. There are no efficient vaccination procedures available and current therapies fail with a large group of patients. Thus, unraveling the molecular basis of the viral replication process is important to understand the mechanisms of viral pathogenesis and to improve the treatment of hepatitis C. Our proposal aims at identifying and functionally characterizing cellular proteins, which, besides hitherto known `host factors¿, support the replication of the HCV RNA genome in the cell¿s cytoplasm and to achieve further insights into the physical organization of the viral replication complex (RC). Recent studies revealed the existence of a high molecular weight cytoplasmic complex that contains HCV host factors such as the hVAP-A protein and the so-called NFAR proteins as well as various undetermined proteins. Considering this complex a likely intermediate of the viral RC it will be purified and its protein components identified by mass-spec analysis. Newly recognized proteins will be tested in terms of a potential role in the intracellular translation and replication cycle of the HCV RNA and the molecular detail of protein-protein interactions analyzed by genetic and biochemical means. Along these lines, we anticipate gaining vital information on the composition of the viral replication machinery and to expand our knowledge on pathogenic features of HCV that are caused by the viral recruitment of cellular components.