Section of Microbial Pathogenesis, Yale University School of Medicine, New Haven, Connecticut, United States of America.
PLoS Pathog. 2011 Oct;7(10):e1002302. doi: 10.1371/journal.ppat.1002302. Epub 2011 Oct 20.
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.
丙型肝炎病毒 (HCV) 核心蛋白被定向到脂滴 (LD) 的表面，这是产生感染性病毒的关键步骤。然而，核心从 LD 招募到新生病毒颗粒的过程尚不清楚。为了研究核心运输的动力学，我们开发了在产生病毒的活细胞中成像功能性核心蛋白的方法。在病毒组装的高峰期，核心在大的、非运动性的 LD 上形成极化帽，与假定的组装部位相邻。此外，还发现 LD 不依赖的、运动性的核心点状结构沿微管运输。重要的是，核心从 LD 招募到这些点状结构，病毒非结构蛋白 NS2 和 NS3-4A 之间的相互作用对这一招募过程至关重要。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作用。
