疱疹病毒组装需要来自细胞核和细胞质膜系统的大分子和膜结构的细胞质缔合。人类巨细胞病毒(HCMV)在其组织核周细胞质病毒组装室(cVAC)的细胞中的研究结果表明,负端定向微管运动有明确的要求,动力蛋白,用于病毒组装。相比之下,单纯疱疹病毒-1(HSV-1)在上皮细胞中的组装,在那里它形成多重分散,外周组装位点仅被微管解聚剂轻度抑制,诺考达唑.这里,我们利用神经元细胞系系统,其中HSV-1形成单个cVAC,并显示动力蛋白及其辅因子动力蛋白定位于cVAC,和动态蛋白与含有病毒体被膜蛋白pUL11的膜相关。我们还表明,病毒膜相关结构蛋白pUL51和病毒包膜糖蛋白gE通过不同的途径到达cVAC。具体来说,gE从质膜取回后到达cVAC,这表明需要一个完整的逆行运输系统。最后,我们证明,在感染的细胞质组装阶段,动态蛋白功能的抑制极大地抑制了cVAC的形成和病毒的产生。重要意义许多病毒重组细胞质膜系统和大分子转运系统以促进后代病毒体的产生。阐明它们实现这一目标的机制可能揭示新的治疗策略,并说明对正常细胞组织至关重要的机制。这里,我们探讨了HSV-1在感染细胞中移动大分子和膜货物以产生病毒组装区室的机制。我们发现这种病毒利用了一个很好的特征,基于微管的运输系统,对破坏微管的药物稳定。
Herpesvirus assembly requires the cytoplasmic association of large macromolecular and membrane structures that derive from both the nucleus and cytoplasmic membrane systems. Results from the study of human cytomegalovirus (HCMV) in cells where it organizes a perinuclear cytoplasmic virus assembly compartment (cVAC) show a clear requirement for the minus-end-directed microtubule motor, dynein, for virus assembly. In contrast, the assembly of herpes simplex virus -1 (HSV-1) in epithelial cells where it forms multiple dispersed, peripheral assembly sites is only mildly inhibited by the microtubule-depolymerizing agent, nocodazole. Here, we make use of a neuronal cell line system in which HSV-1 forms a single cVAC and show that dynein and its co-factor
dynactin localize to the cVAC, and
dynactin is associated with membranes that contain the virion tegument protein pUL11. We also show that the virus membrane-associated structural proteins pUL51 and the viral envelope glycoprotein gE arrive at the cVAC by different routes. Specifically, gE arrives at the cVAC after retrieval from the plasma membrane, suggesting the need for an intact retrograde transport system. Finally, we demonstrate that inhibition of
dynactin function profoundly inhibits cVAC formation and virus production during the cytoplasmic assembly phase of infection.IMPORTANCEMany viruses reorganize cytoplasmic membrane systems and macromolecular transport systems to promote the production of progeny virions. Clarifying the mechanisms by which they accomplish this may reveal novel therapeutic strategies and illustrate mechanisms that are critical for normal cellular organization. Here, we explore the mechanism by which HSV-1 moves macromolecular and membrane cargo to generate a virus assembly compartment in the infected cell. We find that the virus makes use of a well-characterized, microtubule-based transport system that is stabilized against drugs that disrupt microtubules.