Polymerase-tagged respiratory syncytial virus reveals a dynamic rearrangement of the ribonucleocapsid complex during infection.
Polymerase-tagged respiratory syncytial virus reveals a dynamic rearrangement of the ribonucleocapsid complex during infection.
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DOI：
10.1371/journal.ppat.1008987
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发表时间：
2020-10
期刊：
影响因子：
6.7
通讯作者：
Santangelo PJ
中科院分区：
文献类型：
--
作者：
Blanchard EL;Braun MR;Lifland AW;Ludeke B;Noton SL;Vanover D;Zurla C;Fearns R;Santangelo PJ
呼吸道合胞病毒 (RSV) 的核糖核衣壳复合物负责感染期间的病毒 mRNA 转录和病毒复制，但人们对如何实现这种双重功能知之甚少。在这里，我们报告了使用带有 FLAG 标签的大聚合酶蛋白 L 的重组 RSV 病毒来表征和定位病毒感染早期和晚期阶段的 RSV 核糖核衣壳结构。通过邻近连接测定和超分辨率显微镜，揭示了核糖核衣壳复合物中的病毒RNA和蛋白质随着时间的推移，特别是感染后6至8小时之间动态重排，这表明核糖核衣壳结构与其功能之间存在联系。核糖核衣壳重排的时间与 RSV 基因组 RNA 积累的增加相对应，表明这种重排可能与 RNA 复制和二次转录的开始有关。此外，体外转录 mRNA 的 RSV M2-2 早期过表达被证明可以通过重新排列核糖核衣壳复合物来抑制病毒感染。总的来说，这些结果详细说明了对 RSV L 和核糖核衣壳复合物在 RSV 感染期间的定位和活性的重要理解。呼吸道合胞病毒是五岁以下儿童常见的呼吸道疾病。每年，大约有三百万受感染的儿童需要住院治疗。然而，尚无针对呼吸道合胞病毒的疫苗或有效的抗病毒治疗方法。了解呼吸道合胞病毒如何在细胞中复制将为制定更有效的对策提供关键知识。在这里，我们研究了呼吸道合胞病毒聚合酶的大蛋白亚基（负责病毒 mRNA 的合成和复制）与感染过程中其他病毒蛋白之间的相互作用，以及大蛋白亚基随时间在关键病毒结构中的定位。我们的研究为呼吸道合胞病毒聚合酶的定位和活性提供了新的见解，并表明核糖核衣壳复合物重排在呼吸道合胞病毒聚合酶在病毒复制中的功能中起着关键作用，从而更好地了解呼吸道合胞病毒生物学。
The ribonucleocapsid complex of respiratory syncytial virus (RSV) is responsible for both viral mRNA transcription and viral replication during infection, though little is known about how this dual function is achieved. Here, we report the use of a recombinant RSV virus with a FLAG-tagged large polymerase protein, L, to characterize and localize RSV ribonucleocapsid structures during the early and late stages of viral infection. Through proximity ligation assays and super-resolution microscopy, viral RNA and proteins in the ribonucleocapsid complex were revealed to dynamically rearrange over time, particularly between 6 and 8 hours post infection, suggesting a connection between the ribonucleocapsid structure and its function. The timing of ribonucleocapsid rearrangement corresponded with an increase in RSV genome RNA accumulation, indicating that this rearrangement is likely involved with the onset of RNA replication and secondary transcription. Additionally, early overexpression of RSV M2-2 from in vitro transcribed mRNA was shown to inhibit virus infection by rearranging the ribonucleocapsid complex. Collectively, these results detail a critical understanding into the localization and activity of RSV L and the ribonucleocapsid complex during RSV infection. Respiratory syncytial virus is a common respiratory disease among children under the age of five. Yearly, approximately three million children infected require hospitalization for treatment. However, no vaccine or effective antiviral treatment for respiratory syncytial virus yet exists. Understanding how respiratory syncytial virus replicates in cells will provide key knowledge for the development of more effective countermeasures. Here, we investigated the interactions between the large protein subunit of the respiratory syncytial virus polymerase, which is responsible for viral mRNA synthesis and replication, and other viral proteins during infection, as well as the localization of the large protein subunit over time in key viral structures. Our study provides new insights into the localization and activity of the respiratory syncytial virus polymerase, and suggests ribonucleocapsid complex rearrangement plays a key role in the function of the respiratory syncytial virus polymerase in replication of the virus, leading to a better understanding of respiratory syncytial virus biology.
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影响因子：
11.2
作者：
Blanchard, Emmeline L.;Argyropoulou, Danae;Santangelo, Philip J.
通讯作者：
Santangelo, Philip J.
DOI：
10.1099/vir.0.012047-0
发表时间：
2009-09
期刊：
The Journal of general virology
影响因子：
--
作者：
Carlos TS;Young DF;Schneider M;Simas JP;Randall RE
通讯作者：
Randall RE
影响因子：
3.7
作者：
Fearns, R;Peeples, ME;Collins, PL
通讯作者：
Collins, PL
影响因子：
5.4
作者：
EGELMAN, EH;WU, SS;MURTI, G
通讯作者：
MURTI, G
影响因子：
5.6
作者：
Bhella, D;Ralph, A;Yeo, RP
通讯作者：
Yeo, RP