Molecular determinants of hepatitis C virus assembly

 描述(申请人提供)：尽管最近在抗病毒药物开发方面取得了进展，丙型肝炎病毒(丙型肝炎病毒)仍然是慢性肝病和癌症的主要原因，获得治疗的机会有限，耐药性迅速演变，并且没有有效的疫苗。因此，有一个尚未得到满足的需求，即改进治疗和创造疫苗。由于大多数抗病毒疫苗都能诱导针对病毒颗粒的体液反应，因此需要进一步研究以了解丙型肝炎病毒颗粒的结构和组装，这些颗粒通过与血清脂蛋白的一种尚不明确的相互作用而免受抗体中和。丙型肝炎病毒也是一个有价值的模式系统，以了解黄病毒-一个大家族的临床重要的病毒病原体-以及更广泛的正链RNA病毒。一个关键的悬而未决的问题是“正链RNA病毒编码的解旋酶是如何促进病毒组装的？”这项建议建立在我们在前一个资金阶段研究丙型肝炎病毒颗粒组装的成功的基础上，作为一个独特的病毒结构范例和一个了解解旋酶功能的模型系统。通过利用我们以前的成功和采用新的方法，我们将实现以下三个目标：1)确定丙型肝炎病毒解旋酶结构域在病毒组装中的作用；(2)确定病毒组装复合体中的功能相互作用；以及(3)确定载脂蛋白E在病毒结构中的作用。该项目的成功完成将为在深层机制水平上理解病毒解旋酶在被包裹的正链RNA病毒组装过程中的作用奠定基础，为核衣壳组装的协调提供见解，并首次严格测试丙型肝炎病毒颗粒结构的流行模型。

 DESCRIPTION (provided by applicant): Despite recent advances in antiviral drug development, hepatitis C virus (HCV) remains a major cause of chronic liver disease and cancer, access to therapy is limited, drug resistance evolves rapidly, and there is no effective vaccine. Thus, there is an unmet need to improve therapy and to create a vaccine. As most antiviral vaccines elicit humoral responses against virus particles, further research is needed to understand the structure and assembly of HCV particles, which are protected from antibody neutralization through an as-yet-uncharacterized interaction with serum lipoproteins. HCV is also a valuable model system for understanding the Flaviviridae - a large family of clinically important viral pathogens - as well as positive-strand RNA viruses more generally. A key unanswered question is "how do positive-strand RNA virus-encoded helicases contribute to virus assembly?" This proposal builds upon our success in the previous funding period to examine HCV particle assembly, both as a unique paradigm of virus structure and as a model system for understanding helicase function. By leveraging our prior success and adapting new approaches, we will achieve the following three Aims: 1) establish the role of the HCV helicase domain in virus assembly; (2) define functional interactions within the virus assembly complex; and (3) determine the role of Apolipoprotein E in virus structure. Successful completion of this Project will lay the groundwork for understanding, at a deep mechanistic level, the role of viral helicases during the assembly of enveloped, positive-strand RNA viruses, provide insights into the coordination of nucleocapsid assembly, and - for the first time - rigorously test the prevailing model of HCV particle structure.