Mechanisms of HCV Assembly

 描述(申请人提供)：目前，全球近2亿人口[包括美国的400万人]慢性感染丙型肝炎病毒。丙型肝炎病毒会导致严重的肝病，包括慢性肝炎、肝硬变和肝细胞癌。丙型肝炎病毒组装是丙型肝炎病毒复制过程中最不为人所知的步骤，因此，在开发抗丙型肝炎病毒治疗药物以阻断病毒复制这一关键步骤方面进展甚微。这项提案的目标是定义管理丙型肝炎病毒组装过程的调控机制。我们相信，通过开展拟议的研究来实现这一目标，不仅将促进对丙型肝炎病毒组装机制的了解，还将有助于针对丙型肝炎病毒组装步骤的抗病毒药物的发展。根据最近的文献和我们的初步研究，我们提出了一个新的假设，即自然的次优E2-p7加工通过调节依赖于p7的NS2，从而调节依赖于NS2的E2定位到DRM上的病毒组装位点，是导致丙型肝炎病毒组装延迟发生的关键。我们计划通过研究以下特定目标来验证这一假设：特定目标1将阐明NS2亚细胞定位的决定因素。具体目标2将阐明NS2依赖的E2定位于DRM的机制。特异性目标3将确定E2定位在丙型肝炎病毒形态发生的DRM中的意义。实现这些目标将使我们能够定义病毒组装因子靶向过程中涉及的丙型肝炎病毒组装机制的新方面，并有可能建立ER-DRM作为丙型肝炎病毒颗粒萌发的位置。

 DESCRIPTION (provided by applicant): Currently, near 200 million of the world's population [including 4 million persons in the US] is chronically infected with hepatitis C virus (HCV). HCV causes severe liver diseases, including chronic hepatitis, cirrhosis, and hepatocellular carcinoma. HCV assembly is the least understood step during the HCV replication process and, due to this, little progress has been made toward developing the anti-HCV therapeutics to block this critical step in viral replication. The goal of this proposal is to define the regulatory mechanisms governing HCV assembly process. We believe that achieving this goal by performing the proposed studies will not only advance the knowledge of the HCV assembly mechanisms but also help the development of the antivirals targeting HCV assembly step. Based on the recent literature and our preliminary studies, we propose a novel hypothesis that naturally suboptimal E2-p7 processing is critical for the late onset of HCV assembly by regulating p7-dependent NS2, and consequently NS2-dependent E2 localization, to the virus assembly sites at the DRM. We plan to test this hypothesis by investigating the following specific aims: Specific aim 1 will elucidate the determinants of temporal subcellular localization of NS2. Specific aim 2 will elucidate the mechanism of NS2-dependnet E2 localization to the DRM. Specific aim 3 will determine the significance of E2 localization to the DRM on HCV morphogenesis. Accomplishing these aims will allow us to define novel aspects of the HCV assembly mechanisms involved in the virus assembly-factor targeting processes and potentially to establish the ER-DRM as the site of HCV particle budding.