Migration of vesicular stomatitis virus glycoprotein to the nucleus of infected cells
Migration of vesicular stomatitis virus glycoprotein to the nucleus of infected cells
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DOI：
10.1073/pnas.93.16.8268
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发表时间：
1996-08-06
影响因子：
11.1
通讯作者：
Silva, JL
中科院分区：
文献类型：
--
作者：
DaPoian, AT;Gomes, AM;Silva, JL
报道了一种通过病毒蛋白的选择性荧光标记结合数字成像显微镜直接可视化病毒感染早期事件的新方法，病毒感染的早期阶段对于了解病毒复制和发病机制具有重要意义。水疱性口炎病毒是研究最深入的弹状病毒，由负义RNA基因组、五种病毒蛋白和源自宿主细胞的膜脂组成。用异硫氰酸荧光素标记水疱性口炎病毒的糖蛋白，并与标记的病毒一起孵育。小仓鼠肾细胞，感染开始后，首先在细胞内的内吞囊泡中看到标记糖蛋白的荧光。荧光逐渐迁移到受感染细胞的细胞核。感染1小时后，病毒糖蛋白浓缩在细胞核中，并可以在纯化细胞核制剂中完整回收。这些结果表明，病毒 RNA 的脱壳发生在核膜附近，这将先于前导 RNA 的转录进入细胞核，从而关闭细胞 RNA 合成和 DNA 复制。
A new means of direct visualization of the early events of viral infection by selective fluorescence labeling of viral proteins coupled with digital imaging microscopy is reported, The early phases of viral infection have great importance for understanding viral replication and pathogenesis. Vesicular stomatitis virus, the best-studied rhabdovirus, is composed of an RNA genome of negative sense, five viral proteins, and membrane lipids derived from the host cell. The glycoprotein of vesicular stomatitis virus was labeled with fluorescein isothiocyanate, and the labeled virus was incubated with. baby hamster kidney cells, After initiation of infection, the fluorescence of the labeled glycoprotein was first seen inside the cells in endocytic vesicles. The fluorescence progressively migrated to the nucleus of infected cells. After 1 h of infection, the virus glycoprotein was concentrated in the nucleus and could be recovered intact in a preparation of purified nuclei. These results suggest that uncoating of the viral RNA occurs close to the nuclear membrane, which would precede transcription of the leader RNA that enters the nucleus to shut off cellular RNA synthesis and DNA replication.