单纯疱疹病毒1(HSV-1)是一种α疱疹病毒,感染了世界上大多数人口。参与HSV-1颗粒的细胞内转运和胞吐的机制和细胞宿主因子尚未完全了解。为了阐明复制周期中的这些后期步骤,我们开发了HSV-1病毒粒子细胞内运输和胞吐作用的活细胞荧光显微镜检测。这种方法使我们能够跟踪单个病毒颗粒,并使用pH敏感的报道分子确定颗粒胞吐的精确时刻和位置。我们证明HSV-1在外出期间使用宿主细胞的高尔基体后分泌途径。小的GTPase,Rab6与跨高尔基网络的新生分泌囊泡结合,发挥重要作用,但非必要的,在质膜的囊泡运输和胞吐中的作用,因此使其成为高尔基体和后高尔基体分泌途径的有用标记。我们表明HSV-1粒子与Rab6a在高尔基体区域共定位,将Rab6a与细胞外围共交,并从Rab6a囊泡中胞吐。与以前的报告一致,我们发现HSV-1颗粒在受感染细胞的优先出口位点积累。分泌途径介导这种优先/极化的出口,因为Rab6a囊泡在未感染细胞中类似地积累在质膜附近。这些数据表明,在粒子包络之后,HSV-1的出口遵循预先存在的细胞分泌途径退出感染的细胞,而不是新的,病毒诱导的机制。
目的:单纯疱疹病毒1型(HSV-1)感染大多数人。它会建立终身潜伏感染,偶尔会重新激活,通常引起特征性口腔或生殖器病变。很少在健康的自然宿主中,但更常见于人畜共患感染和老年人,新生,或者免疫功能低下的患者,HSV-1可引起严重的疱疹性脑炎。HSV-1使用的精确细胞机制仍然是一个重要的研究领域。特别是,新组装的病毒颗粒用于从感染细胞中退出的出口途径尚不清楚.在这项研究中,我们使用荧光显微镜观察从细胞中排出的单个病毒颗粒,发现HSV-1颗粒利用了预先存在的细胞分泌途径.
Herpes simplex virus 1 (HSV-1) is an alpha herpesvirus that infects a majority of the world population. The mechanisms and cellular host factors involved in the intracellular transport and exocytosis of HSV-1 particles are not fully understood. To elucidate these late steps in the replication cycle, we developed a live-cell fluorescence microscopy assay of HSV-1 virion intracellular trafficking and exocytosis. This method allows us to track individual virus particles and identify the precise moment and location of particle exocytosis using a pH-sensitive reporter. We show that HSV-1 uses the host cell\'s post-Golgi secretory pathway during egress. The small GTPase, Rab6, binds to nascent secretory vesicles at the trans-Golgi network and plays important, but non-essential, roles in vesicle traffic and exocytosis at the plasma membrane, therefore making it a useful marker of the Golgi and post-Golgi secretory pathway. We show that HSV-1 particles colocalize with Rab6a in the region of the Golgi, cotraffic with Rab6a to the cell periphery, and undergo exocytosis from Rab6a vesicles. Consistent with previous reports, we find that HSV-1 particles accumulate at preferential egress sites in infected cells. The secretory pathway mediates this preferential/polarized egress, since Rab6a vesicles accumulate near the plasma membrane similarly in uninfected cells. These data suggest that, following particle envelopment, HSV-1 egress follows a pre-existing cellular secretory pathway to exit infected cells rather than novel, virus-induced mechanisms.
OBJECTIVE: Herpes simplex virus 1 (HSV-1) infects a majority of people. It establishes a life-long latent infection and occasionally reactivates, typically causing characteristic oral or genital lesions. Rarely in healthy natural hosts, but more commonly in zoonotic infections and in elderly, newborn, or immunocompromised patients, HSV-1 can cause severe herpes encephalitis. The precise cellular mechanisms used by HSV-1 remain an important area of research. In particular, the egress pathways that newly assembled virus particles use to exit from infected cells are unclear. In this study, we used fluorescence microscopy to visualize individual virus particles exiting from cells and found that HSV-1 particles use the pre-existing cellular secretory pathway.