Members of the Reoviridae family assemble virus factories within the cytoplasm of infected cells to replicate and assemble virus particles. Bluetongue virus (BTV) forms virus inclusion bodies (VIBs) that are aggregates of viral RNA, certain viral proteins, and host factors, and have been shown to be sites of the initial assembly of transcriptionally active virus-like particles. This study sought to characterize the formation, composition, and ultrastructure of VIBs, particularly in relation to virus replication. In this study we have utilized various microscopic techniques, including structured illumination microscopy, and virological assays to show for the first time that the outer capsid protein VP5, which is essential for virus maturation, is also associated with VIBs. The addition of VP5 to assembled virus cores exiting VIBs is required to arrest transcriptionally active core particles, facilitating virus maturation. Furthermore, we observed a time-dependent association of the glycosylated non-structural protein 3 (NS3) with VIBs, and report on the importance of the two polybasic motifs within NS3 that facilitate virus trafficking and egress from infected cells at the plasma membrane. Thus, the presence of VP5 and the dynamic nature of NS3 association with VIBs that we report here provide novel insight into these previously less well-characterized processes.

中文翻译：

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蓝舌病毒复制周期中结构蛋白和非结构蛋白的差异化定位。

呼肠孤病毒科成员在感染细胞的细胞质内组装病毒工厂，以复制和组装病毒颗粒。蓝舌病病毒（BTV）形成病毒包涵体（VIB），它们是病毒RNA，某些病毒蛋白和宿主因子的聚集体，并已被证明是转录活性病毒样颗粒的初始装配位点。这项研究试图表征VIB的形成，组成和超微结构，特别是在病毒复制方面。在这项研究中，我们利用了各种显微技术，包括结构照明显微镜和病毒学检测，首次证明了病毒成熟的外衣壳蛋白VP5也与VIBs有关。需要在退出VIB的组装病毒核心中添加VP5，以阻止转录活性核心颗粒，从而促进病毒成熟。此外，我们观察到糖基化非结构蛋白3（NS3）与VIB的时间依赖性关联，并报告了NS3中两个多基元基序的重要性，这些基元有助于病毒的运输和从被感染细胞的质膜流出。因此，我们在此报告的VP5的存在和NS3与VIB的动态关联为我们提供了对这些以前不太好的过程的新颖见解。并报道了NS3中两个多基元基序的重要性，这些基元有助于病毒的运输和从质膜上被感染的细胞中逸出。因此，我们在此报告的VP5的存在以及NS3与VIB关联的动态性质为这些以前不太为人所知的过程提供了新颖的见解。并报道了NS3中两个多基元基序的重要性，这些基元有助于病毒的运输和从质膜上被感染的细胞中逸出。因此，我们在此报告的VP5的存在以及NS3与VIB关联的动态性质为这些以前不太为人所知的过程提供了新颖的见解。