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。目前还没有有效的疫苗接种程序，目前的治疗方法对大量患者无效。因此，揭示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。我们的建议旨在鉴定和功能表征细胞蛋白，这些蛋白除了已知的“宿主因子”外，还支持HCV RNA基因组在细胞细胞质中的复制，并进一步了解病毒复制复合体（RC）的物理组织。最近的研究表明，存在一种高分子量的细胞质复合体，其中包含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质。考虑到该复合物可能是病毒RC的中间产物，将对其进行纯化，并通过质谱分析鉴定其蛋白质成分。新识别的蛋白质将在HCV RNA的细胞内翻译和复制周期中的潜在作用方面进行测试，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。沿着这些思路，我们期望获得关于病毒复制机制组成的重要信息，并扩大我们对由病毒募集细胞成分引起的HCV致病特征的认识。

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.