Characterization of herpes simplex virus-containing organelles by subcellular fractionation: Role for organelle acidification in assembly of infectious particles
Characterization of herpes simplex virus-containing organelles by subcellular fractionation: Role for organelle acidification in assembly of infectious particles
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DOI：
10.1128/jvi.75.3.1236-1251.2001
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发表时间：
2001-02-01
影响因子：
5.4
通讯作者：
Wilson, DW
中科院分区：
文献类型：
--
作者：
Harley, CA;Dasgupta, A;Wilson, DW
单纯疱疹病毒（HSV）胞浆区室的定义尚不清楚。目前还不清楚哪些细胞器包含大部分的贩运，病毒粒子和病毒粒子占据的生产，而不是有缺陷的装配途径。这些问题由于HSV感染的细胞在数小时内持续产生病毒而变得更加复杂。因此，病毒组装和输出的所有阶段共存，使得不可能确定事件的顺序及其动力学。为了解决这些问题，我们已经建立了检测方法来监测衣壳和病毒粒子在细胞提取物中的存在，并从正常感染的细胞和经历单一同步病毒流出波的细胞中制备含有HSV的细胞器。我们发现，在这两种情况下，HSV粒子退出细胞核和积累的细胞器，cofraceousness与trans-Golgi网络（TGN）和内涵体。除了在它们的管腔中携带有包膜的感染性病毒体之外，携带HSV的细胞器还显示附着于它们的细胞质表面的无包膜衣壳。氯喹或巴弗洛霉素A的细胞器pH值的中和导致非感染性包膜颗粒的积累。我们的结论是TGN/内吞网络的细胞器在感染性HSV的组装和运输中发挥了重要作用。
The cytoplasmic compartments occupied by exocytosing herpes simplex virus (HSV) are poorly defined. It is unclear which organelles contain the majority of trafficking,virions and which are occupied by virions on a productive rather than defective assembly pathway. These problems are compounded by the bet that HSV-infected cells produce virus continuously over many hours. All stages in viral assembly and export therefore coexist, making it impossible to determine the sequence of events and their kinetics. To address these problems, we have established assays to monitor the presence of capsids and em eloped virions in cell extracts and prepared HSV-containing organelles from normally infected cells and from cells undergoing a single synchronized wave of viral egress. We find that, in both cases, HSV particles exit the nucleus and accumulate in organelles which cofractionate with the trans-Golgi network (TGN) and endosomes. In addition to carrying enveloped infectious virions in their lumen, HSV-bearing organelles also displayed nonenveloped capsids attached to their cytoplasmic surface. Neutralization of organellar pH by chloroquine or bafilomycin A resulted in the accumulation of noninfectious enveloped particles. We conclude that the organelles of the TGN/endocytic network play a hey role in the assembly and trafficking of infectious HSV.