Molecular determinants of hepatitis C virus assembly

 描述（由申请人提供）：尽管最近在抗病毒药物开发方面取得了进展，但丙型肝炎病毒（HCV）仍然是慢性肝病和癌症的主要原因，获得治疗的机会有限，耐药性迅速发展，并且没有有效的疫苗。因此，存在改进治疗和产生疫苗的未满足的需求。由于大多数抗病毒疫苗引起针对病毒颗粒的体液应答，因此需要进一步研究以了解HCV颗粒的结构和组装，HCV颗粒通过与血清脂蛋白的尚未表征的相互作用而免受抗体中和。HCV也是一个有价值的模型系统，用于了解黄病毒科-一个临床上重要的病毒病原体的大家族-以及更普遍的正链RNA病毒。一个关键的未解问题是“正链RNA病毒编码的解旋酶如何有助于病毒组装？“这项提案建立在我们在前一个资助期成功研究HCV颗粒组装的基础上，既作为病毒结构的独特范例，又作为理解解旋酶功能的模型系统。通过利用我们先前的成功和采用新的方法，我们将实现以下三个目标：1）建立HCV解旋酶结构域在病毒组装中的作用;（2）定义病毒组装复合物内的功能相互作用;（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理解在深层机制水平上病毒解旋酶在包膜正链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HCV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.