Trafficking of hepatitis C virus core protein during virus particle assembly.
Trafficking of hepatitis C virus core protein during virus particle assembly.
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DOI：
10.1371/journal.ppat.1002302
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发表时间：
2011-10
期刊：
影响因子：
6.7
通讯作者：
Lindenbach BD
中科院分区：
文献类型：
--
作者：
Counihan NA;Rawlinson SM;Lindenbach BD
丙型肝炎病毒(丙型肝炎病毒)的核心蛋白被定向到脂滴(LD)的表面，这是产生传染性病毒的关键步骤。然而，核心如何从LD招募到新生的病毒颗粒中的过程还不是很清楚。为了研究核心转运的动力学，我们开发了在活的、产生病毒的细胞中对功能核心蛋白进行成像的方法。在病毒组装的高峰期，核心在大的、静止的LD上形成极化的帽子，靠近假定的组装位置。此外，还发现非LD依赖的、可移动的核心点沿微管运输。重要的是，CORE是从LDS招募到这些斑点中的，病毒NS2和NS3-4A蛋白之间的相互作用在这个招募过程中是必不可少的。这些数据揭示了核心运输的新方面，并确定了病毒非结构蛋白在病毒颗粒组装中的新角色。丙型肝炎病毒(丙型肝炎病毒)感染全球近2亿人，导致急性和慢性肝病。尽管存在一些抗病毒治疗，但它们并不是对所有的丙型肝炎病毒都完全有效，而且有严重的副作用。为了开发更有效的治疗策略，需要更好地了解丙型肝炎病毒如何在感染细胞中复制。在我们的研究中，我们开发了一种方法，通过荧光标记病毒的结构成分核心蛋白来可视化丙型肝炎病毒颗粒组装的早期步骤。在蛋白质翻译后不久，核心就被运送到靠近病毒组装部位的大而不动的脂滴的表面。核心也可观察到沿微管移动的极易移动的点。通过使用病毒组装的抑制剂和组装缺陷的病毒突变体，我们发现核心是从脂滴中招募到这些斑点中的，这一过程是由丙型肝炎病毒非结构蛋白的相互作用介导的。我们的工作描述了研究核心蛋白在感染细胞中运输的新方法，使我们能够更好地定义传染性丙型肝炎病毒颗粒组装的各个方面。
Hepatitis C virus (HCV) core protein is directed to the surface of lipid droplets (LD), a step that is essential for infectious virus production. However, the process by which core is recruited from LD into nascent virus particles is not well understood. To investigate the kinetics of core trafficking, we developed methods to image functional core protein in live, virus-producing cells. During the peak of virus assembly, core formed polarized caps on large, immotile LDs, adjacent to putative sites of assembly. In addition, LD-independent, motile puncta of core were found to traffic along microtubules. Importantly, core was recruited from LDs into these puncta, and interaction between the viral NS2 and NS3-4A proteins was essential for this recruitment process. These data reveal new aspects of core trafficking and identify a novel role for viral nonstructural proteins in virus particle assembly. Hepatitis C virus (HCV) infects almost 200 million people worldwide, causing both acute and chronic liver disease. Although some antiviral treatments exist, they are not fully effective against all HCV genotypes and have serious side effects. In order to develop more effective treatment strategies, a better understanding of how HCV replicates in infected cells is required. In our study, we developed methods to visualize early steps in HCV particle assembly by fluorescently labeling core protein, a structural component of the virus. Soon after protein translation, core trafficked to the surface of large, immobile lipid droplets that were adjacent to sites of virus assembly. Core was also observed in highly motile puncta that traveled along microtubules. By using inhibitors of virus assembly and assembly-deficient viral mutants, we showed that core is recruited from lipid droplets into these puncta, and that this process was mediated by the interaction of HCV nonstructural proteins. Our work describes new methods to study the trafficking of core protein in infected cells, allowing us to better define aspects of infectious HCV particle assembly.
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