Apolipoprotein E Interacts with Hepatitis C Virus Nonstructural Protein 5A and Determines Assembly of Infectious Particles
Apolipoprotein E Interacts with Hepatitis C Virus Nonstructural Protein 5A and Determines Assembly of Infectious Particles
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DOI：
10.1002/hep.23278
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发表时间：
2010-01-01
期刊：
影响因子：
13.5
通讯作者：
Schuster, Catherine
中科院分区：
文献类型：
--
作者：
Benga, Wagane J. A.;Krieger, Sophie E.;Schuster, Catherine
慢性丙型肝炎病毒（HCV）感染是世界范围内肝脏疾病的主要原因。限制丙型肝炎病毒感染人肝细胞表明肝脏特异性宿主因子在病毒生命周期中发挥作用。使用酵母-双杂交系统，我们鉴定了载脂蛋白E （apoE）作为肝脏来源的宿主因子特异性地与HCV非结构性相互作用。蛋白5A (NS5A)，但不与其他病毒蛋白结合。apoE和NS5A在感染性HCV细胞培养模型系统中的共免疫沉淀和共定位研究证实了apoE-NS5A相互作用与病毒感染的相关性。沉默apoE表达可显著抑制感染性颗粒的产生，但不影响病毒的进入和复制。对apoE表达沉默的肝源性细胞颗粒产生的分析显示，感染性颗粒的组装和释放受到损害。装配缺陷病毒突变体中apoE-NS5A结合缺失或减少，支持了apoE-NS5A相互作用与病毒颗粒产生的功能相关性。结论：这些结果提示NS5A募集apoE对病毒的组装和感染性病毒颗粒的释放具有重要意义。这些发现对于理解HCV生命周期和开发针对HCV-脂蛋白相互作用的新型抗病毒策略具有重要意义。(肝脏病学2010;51:43-53)。
Chronic hepatitis C virus (HCV) infection is a major cause of liver disease worldwide. Restriction of HCV infection to human hepatocytes suggests that liver-specific host factors play a role in the viral life cycle. Using a yeast-two-hybrid system, we identified apolipoprotein E (apoE) as a liver-derived host factor specifically interacting with HCV nonstructural. protein 5A (NS5A) but not with other viral proteins. The relevance of apoE-NS5A interaction for viral infection was confirmed by co-immunoprecipitation and co-localization studies of apoE and NS5A in an infectious HCV cell culture model system. Silencing apoE expression resulted in marked inhibition of infectious particle production without affecting viral entry and replication. Analysis of particle production in liver-derived cells with silenced apoE expression showed impairment of infectious particle assembly and release. The functional relevance of the apoE-NS5A interaction for production of viral particles was supported by loss or decrease of apoE-NS5A binding in assembly-defective viral mutants. Conclusion: These results suggest that recruitment of apoE by NS5A is important for viral assembly and release of infectious viral particles. These findings have important implications for understanding the HCV life cycle and the development of novel antiviral strategies targeting HCV-lipoprotein interaction. (HEPATOLOGY 2010;51:43-53.)