Aquareovirus NS80 Initiates Efficient Viral Replication by Retaining Core Proteins within Replication-Associated Viral Inclusion Bodies.
Aquareovirus NS80 Initiates Efficient Viral Replication by Retaining Core Proteins within Replication-Associated Viral Inclusion Bodies.
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Aquareovirus NS80 通过在复制相关的病毒包涵体内保留核心蛋白来启动有效的病毒复制
DOI：
10.1371/journal.pone.0126127
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发表时间：
2015
期刊：
影响因子：
3.7
通讯作者：
Fang Q
中科院分区：
文献类型：
--
作者：
Yan L;Zhang J;Guo H;Yan S;Chen Q;Zhang F;Fang Q
病毒包涵体（viral inclusion bodies，VIBs）是呼肠孤病毒复制和组装的特异性细胞内区室。在我们以前的研究中，已经确定了水生呼肠孤病毒非结构蛋白NS 80是形成球形VIBs的主要成分。在这项研究中，我们研究了NS 80在病毒结构蛋白表达和病毒复制中的作用。免疫荧光分析表明，NS 80可以保留五个核心蛋白或内衣壳蛋白（VP 1-VP 4和VP 6），但没有外衣壳蛋白（VP 5和VP 7），在共转染或感染细胞的VIBs。进一步的免疫共沉淀分析证实，NS 80可以分别与每个核心蛋白相互作用。此外，我们发现新合成的病毒RNA与VIBs共定位。对病毒结构蛋白表达的时程分析表明，病毒结构蛋白NS 80的表达最先被检测到，其次是内壳蛋白VP 3，最后才是其他内壳蛋白的表达，表明VIBs对病毒核心框架或子代病毒粒子的形成是必不可少的。此外，通过shRNA敲低NS 80不仅抑制了水生呼肠孤病毒结构蛋白的表达，而且抑制了病毒感染。这些结果表明，NS 80为基础的VIBs在感染的早期就形成了，并且NS 80能够协调病毒结构蛋白的表达和病毒的复制。
Viral inclusion bodies (VIBs) are specific intracellular compartments for reoviruses replication and assembly. Aquareovirus nonstructural protein NS80 has been identified to be the major constituent for forming globular VIBs in our previous study. In this study, we investigated the role of NS80 in viral structural proteins expression and viral replication. Immunofluorescence assays showed that NS80 could retain five core proteins or inner-capsid proteins (VP1-VP4 and VP6), but not outer-capsid proteins (VP5 and VP7), within VIBs in co-transfected or infected cells. Further co-immunoprecipitation analysis confirmed that NS80 could interact with each core protein respectively. In addition, we found that newly synthesized viral RNAs co-localized with VIBs. Furthermore, time-course analysis of viral structural proteins expression showed that the expression of NS80 was detected first, followed by the detection of inner shell protein VP3, and then of other inner-capsid proteins, suggesting that VIBs were essential for the formation of viral core frame or progeny virion. Moreover, knockdown of NS80 by shRNA not only inhibited the expression of aquareovirus structural proteins, but also inhibited viral infection. These results indicated that NS80-based VIBs were formed at earlier stage of infection, and NS80 was able to coordinate the expression of viral structural proteins and viral replication.
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DOI：
10.1073/pnas.50.2.268
发表时间：
1963-01-01
影响因子：
11.1
作者：
DALES, S
通讯作者：
DALES, S
杆状病毒中表达的草鱼呼肠孤病毒（GCRV）的VP2蛋白表现出RNA聚合酶活性
影响因子：
5.5
作者：
Fan, Liming;Liu, Huan;Fang, Qin
通讯作者：
Fang, Qin
影响因子：
6.4
作者：
Desmet EA;Anguish LJ;Parker JS
通讯作者：
Parker JS
影响因子：
3.8
作者：
Eichwald, C;Rodriguez, JF;Burrone, OR
通讯作者：
Burrone, OR
影响因子：
5.4
作者：
Broering, TJ;Kim, J;Parker, JSL
通讯作者：
Parker, JSL