Early stages of influenza virus entry into Mv-1 lung cells: Involvement of dynamin
Early stages of influenza virus entry into Mv-1 lung cells: Involvement of dynamin
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DOI：
10.1006/viro.1999.0109
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发表时间：
2000-02-01
期刊：
影响因子：
3.7
通讯作者：
Whittaker, GR
中科院分区：
文献类型：
--
作者：
Roy, AMM;Parker, JS;Whittaker, GR
病毒通常有两种进入和脱壳机制之一。它们可以通过内吞作用或在质膜上直接融合进入细胞。我们已经建立了一种新的貂肺（MV-1）细胞系，表达的显性干扰形式的发动蛋白-1（K44 A）的控制下的四环素反应元件，并研究了流感感染的早期事件，使用这些细胞。我们发现流感病毒与诱导细胞和未诱导细胞的结合相等，但在表达K44 A的细胞中，电子显微镜显示病毒被困在深包被的凹坑和包含离散包被区域的不规则形状的管状结构中。我们还表明，通过免疫荧光和共聚焦显微镜，传入的病毒进入细胞核被阻止在K44 A表达细胞。免疫荧光显微镜检测病毒复制，并强烈抑制在早期和晚期感染后的K44 A表达细胞。当通过流感空斑试验分析时，在表达K44 A发动蛋白的细胞中，病毒感染性被抑制了约2个对数单位。总的来说，这些数据表明，发动蛋白是需要有效的流感病毒进入，大概是由于其功能的囊泡从包被的坑释放。(C)北京大学出版社.
Viruses generally have one of two mechanisms for entry and uncoating. They can enter the cell either by endocytosis or by direct fusion at the plasma membrane. We have established a novel mink lung (Mv-l) cell line that expresses a dominant-interfering form of dynamin-1 (K44A) under the control of a tetracycline-responsive element and studied the early events in influenza infection using these cells. We found that influenza virus binds equally to both induced and uninduced cells, but in K44A-expressing cells, electron microscopy showed viruses trapped in deep coated pits and irregular-shaped tubular structures that contain discrete coated regions. We also show by immunofluorescence and confocal microscopy that entry of incoming virus into the nucleus is blocked in K44A-expressing cells. Virus replication was assayed by immunofluorescence microscopy and was strongly inhibited at both early and late times postinfection in K44A-expressing cells. Virus infectivity was inhibited by similar to 2 log units in cells expressing K44A dynamin when analyzed by influenza plaque assay. Overall these data show that dynamin is required for efficient influenza virus entry, presumably due to its function in release of vesicles from coated pits. (C) 2000 Academic Press.