RSV glycoprotein and genomic RNA dynamics reveal filament assembly prior to the plasma membrane.
RSV glycoprotein and genomic RNA dynamics reveal filament assembly prior to the plasma membrane.
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DOI：
10.1038/s41467-017-00732-z
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发表时间：
2017-09-22
影响因子：
16.6
通讯作者：
Santangelo PJ
中科院分区：
文献类型：
--
作者：
Vanover D;Smith DV;Blanchard EL;Alonas E;Kirschman JL;Lifland AW;Zurla C;Santangelo PJ
人呼吸道合胞病毒G蛋白在丝状病毒粒子的进入和装配中起重要作用。在这里，我们报告使用荧光标记的大豆凝集素选择性地标记呼吸道合胞病毒G蛋白在活细胞中，而不破坏呼吸道合胞病毒的感染性或丝状体的形成，并允许审问呼吸道合胞病毒病毒粒子组装。使用这种方法，我们发现质膜结合的呼吸道合胞病毒G通过网格蛋白介导的内吞作用迅速从膜上分离。随后动态形成丝状和分支呼吸道合胞病毒颗粒，并在重新插入质膜之前与基因组核糖核蛋白和小窝相关囊泡组装。我们证明，这些过程被中断的微管和抑制分子马达停止。总的来说，我们的研究结果表明，呼吸道合胞病毒装配，病毒丝的产生和基因组RNA加载之前插入质膜。由于缺乏适用于活细胞成像的技术，丝状RSV颗粒的组装不完全理解。Vanover等人使用标记的大豆凝集素选择性地标记RSV G蛋白，并显示丝状RSV装配如何在细胞质中启动，使用从质膜回收的G蛋白。
The human respiratory syncytial virus G protein plays an important role in the entry and assembly of filamentous virions. Here, we report the use of fluorescently labeled soybean agglutinin to selectively label the respiratory syncytial virus G protein in living cells without disrupting respiratory syncytial virus infectivity or filament formation and allowing for interrogations of respiratory syncytial virus virion assembly. Using this approach, we discovered that plasma membrane-bound respiratory syncytial virus G rapidly recycles from the membrane via clathrin-mediated endocytosis. This event is then followed by the dynamic formation of filamentous and branched respiratory syncytial virus particles, and assembly with genomic ribonucleoproteins and caveolae-associated vesicles prior to re-insertion into the plasma membrane. We demonstrate that these processes are halted by the disruption of microtubules and inhibition of molecular motors. Collectively, our results show that for respiratory syncytial virus assembly, viral filaments are produced and loaded with genomic RNA prior to insertion into the plasma membrane. Assembly of filamentous RSV particles is incompletely understood due to a lack of techniques suitable for live-cell imaging. Here Vanover et al. use labeled soybean agglutinin to selectively label RSV G protein and show how filamentous RSV assembly, initiated in the cytoplasm, uses G protein recycled from the plasma membrane.
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影响因子：
5.4
作者：
Kolokoltsov, Andrey A.;Deniger, Drew;Davey, Robert A.
通讯作者：
Davey, Robert A.
DOI：
10.1073/pnas.2434327100
发表时间：
2003-12-09
影响因子：
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作者：
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影响因子：
5.4
作者：
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通讯作者：
Peeples, Mark E.