The N-Terminal of Aquareovirus NS80 Is Required for Interacting with Viral Proteins and Viral Replication.
The N-Terminal of Aquareovirus NS80 Is Required for Interacting with Viral Proteins and Viral Replication.
复制标题
Aquareovirus NS80 的 N 末端是与病毒蛋白相互作用和病毒复制所必需的
DOI：
10.1371/journal.pone.0148550
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发表时间：
2016
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：
Zhang J;Guo H;Chen Q;Zhang F;Fang Q
呼肠孤病毒的复制和组装发生在病毒包涵体内，所述病毒包涵体形成于感染细胞的细胞质的特定细胞内区室中。以往的研究表明，水生呼肠孤病毒NS 80能够形成包涵体，并将病毒蛋白保留在其包涵体中。为了更好地了解NS 80在病毒复制和组装中的作用，本研究对NS 80与其他病毒蛋白在水生呼肠孤病毒复制中的功能区域进行了研究。使用缺失突变分析和基于轮状病毒NSP 5的蛋白关联平台来检测关联区域。免疫荧光显示NS 80的不同N端区域可与病毒蛋白VP 1、VP 4、VP 6和NS 38结合。进一步的免疫共沉淀分析证实了VP 1、VP 4、VP 6和NS 38分别与覆盖NS 80 N-末端氨基酸（aa，1-471）的不同区域之间的相互作用。此外，去除与蛋白VP 1、VP 4、VP 6或NS 38相互作用所需的NS 80 N-末端序列不仅在基于NS 80 shRNA的复制互补测定中阻止了NS 80支持病毒复制的能力，而且还抑制了水生呼肠孤病毒蛋白的表达，表明NS 80的N-末端区域对于病毒复制是必需的。这些结果为进一步了解NS 80在水生呼肠孤病毒感染过程中病毒复制和组装中的作用提供了基础。
Reovirus replication and assembly occurs within viral inclusion bodies that formed in specific intracellular compartments of cytoplasm in infected cells. Previous study indicated that aquareovirus NS80 is able to form inclusion bodies, and also can retain viral proteins within its inclusions. To better understand how NS80 performed in viral replication and assembly, the functional regions of NS80 associated with other viral proteins in aquareovirus replication were investigated in this study. Deletion mutational analysis and rotavirus NSP5-based protein association platform were used to detect association regions. Immunofluorescence images indicated that different N-terminal regions of NS80 could associate with viral proteins VP1, VP4, VP6 and NS38. Further co-immunoprecipitation analysis confirmed the interaction between VP1, VP4, VP6 or NS38 with different regions covering the N-terminal amino acid (aa, 1–471) of NS80, respectively. Moreover, removal of NS80 N-terminal sequences required for interaction with proteins VP1, VP4, VP6 or NS38 not only prevented the capacity of NS80 to support viral replication in NS80 shRNA-based replication complementation assays, but also inhibited the expression of aquareovirus proteins, suggesting that N-terminal regions of NS80 are necessary for viral replication. These results provided a foundational basis for further understanding the role of NS80 in viral replication and assembly during aquareovirus infection.