Immunolocalization of the dengue virus nonstructural glycoprotein NS1 suggests a role in viral RNA replication
Immunolocalization of the dengue virus nonstructural glycoprotein NS1 suggests a role in viral RNA replication
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DOI：
10.1006/viro.1996.0307
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发表时间：
1996-06-01
期刊：
影响因子：
3.7
通讯作者：
Young, YR
中科院分区：
文献类型：
--
作者：
Mackenzie, JM;Jones, MK;Young, YR
虽然许多工作者已经研究了黄病毒非结构糖蛋白NS 1在感染细胞中的成熟和加工，但这些研究对NS 1的可能功能几乎没有提供任何见解。在这项研究中，我们研究了NS 1的亚细胞定位的免疫荧光和肌免疫电镜感染的Vero和C6/36细胞。NS 1与细胞膜紧密结合，尤其是与囊泡包和靶细胞质空泡紧密结合。令人惊讶的是，NS 1不与成熟的病毒颗粒相关联，这一发现与该蛋白在病毒体组装和/或成熟中的假定作用不一致。然而，双标记实验在共聚焦免疫荧光和冷冻免疫EM研究中都揭示了NS 1与病毒dsRNA复制形式的共定位。此外，在感染的Vero和C6/36细胞中分别观察到的dsRNA定位于囊泡包和细胞质空泡，表明这些结构可能包含黄病毒复制复合物。这些发现为NS 1的成熟途径提供了进一步的见解，并表明该蛋白在病毒RNA复制中可能发挥作用。(C)出版社：Academic Press，Inc.
Although many workers have investigated the maturation and processing of the flavivirus nonstructural glycoprotein NS1 in infected cells, these studies have provided little insight into a possible function for NS1. In this study we investigated the subcellular localization of NS1 both by immunofluorescence and Myo-immuno electron microscopy of infected Vero and C6/36 cells. NS1 was found to he tightly associated with intracellular membranes, in particular with Vesicle packets and targe cytoplasmic vacuoles. Surprisingly, NS1 did not associate with mature virus particles, a finding that is inconsistent with the postulated role for this protein in virion assembly and/or maturation. However, dual-labeling experiments did reveal in both the confocal immunofluorescence and cryo-immuno EM studies the colocalization of NS1 with the viral dsRNA replicative form. Furthermore, localization of the dsRNA to the vesicle packets and cytoplasmic vacuoles seen in infected Vero and C6/36 cells, respectively, suggests that these structures may comprise the flavivirus replication complex. These findings provide further insights into the maturation pathway of NS1 and suggest a possible role for this protein in viral RNA replication. (C) 1996 Academic Press, Inc.