Caveolin-1 is incorporated into mature respiratory syncytial virus particles during virus assembly on the surface of virus-infected cells
Caveolin-1 is incorporated into mature respiratory syncytial virus particles during virus assembly on the surface of virus-infected cells
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DOI：
10.1099/0022-1317-83-3-611
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发表时间：
2002-03-01
影响因子：
3.8
通讯作者：
Sugrue, RJ
中科院分区：
文献类型：
--
作者：
Brown, G;Aitken, J;Sugrue, RJ
我们采用免疫荧光显微镜和透射电子显微镜检查呼吸道合胞病毒（RSV）在Vero细胞系C1008中的组装和成熟。RSV在顶端细胞表面以从质膜延伸的丝状形式成熟。我们观察到，包含病毒核糖核蛋白（RNP）核心的包涵体主要出现在质膜下，从那里RSV丝在细胞表面成熟过程中形成。通过共聚焦显微镜比较模拟感染和RSV感染的细胞，发现小窝蛋白-1（cav-1）荧光染色模式发生了显着变化。通过免疫电子显微镜分析表明，RSV细丝形成在细胞表面膜上的cav-1簇附近。此外，免疫电镜显示cav-1与早期出芽的RSV密切相关。通过共聚焦显微镜的进一步分析表明，cav-1随后被掺入宿主细胞膜上成熟的RSV细丝的包膜中，但不与其他病毒结构如病毒RNP相关。尽管cav-1已整合到成熟病毒中，但它定位在簇中，而不是沿着病毒丝的长度均匀分布。此外，当通过免疫阴性染色检查来自感染细胞的组织培养基中的RSV颗粒时，清楚地证明了病毒包膜上存在cav-1。总的来说，这些发现表明cav-1在排出期间被并入成熟RSV颗粒的包膜中。
We have employed immunofluorescence microscopy and transmission electron microscopy to examine the assembly and maturation of respiratory syncytial virus (RSV) in the Vero cell line C1008. RSV matures at the apical cell surface in a filamentous form that extends from the plasma membrane. We observed that inclusion bodies containing viral ribonucleoprotein (RNP) cores predominantly appeared immediately below the plasma membrane, from where RSV filaments form during maturation at the cell surface. A comparison of mock-infected and RSV-infected cells by confocal microscopy revealed a significant change in the pattern of caveolin-1 (cav-1) fluorescence staining. Analysis by immuno-electron microscopy showed that RSV filaments formed in close proximity to cav-1 clusters at the cell surface membrane. In addition, immuno-electron microscopy showed that cav-1 was closely associated with early budding RSV. Further analysis by confocal microscopy showed that cav-1 was subsequently incorporated into the envelope of RSV filaments maturing on the host cell membrane, but was not associated with other virus structures such as the viral RNPs. Although cav-1 was incorporated into the mature virus, it was localized in clusters rather than being uniformly distributed along the length of the viral filaments. Furthermore, when RSV particles in the tissue culture medium from infected cells were examined by immuno-negative staining, the presence of cav-1 on the viral envelope was clearly demonstrated. Collectively, these findings show that cav-1 is incorporated into the envelope of mature RSV particles during egress.