轮状病毒(RV)分为9种,A-D和F-J,物种A是研究最多的。在物种A(RVA)的轮状病毒中,复制发生在病毒质中,它们是由主要构件蛋白NSP5、NSP2和VP2组成的胞浆球状内含物。NSP5与NSP2或VP2在未感染细胞中的共表达导致病毒质样结构(VLSs)的形成。尽管在形态上与病毒质相同,VLSs不产生病毒后代,但可作为研究复杂病毒质的极好工具。由于缺乏特异性抗体和合适的细胞培养系统,关于非RVA病毒质存在知识空白。在这项研究中,我们探索了来自非RVA物种的NSP5和NSP2形成VLS的能力。这两种蛋白质的共表达导致RV物种A中的球状VLS,B,D,F,G,而我,而RVC形成丝状VLS。RV物种H和J的NSP5和NSP2的共表达没有导致VLS形成。有趣的是,所有RV物种的NSP5自低聚,有了有序的C端区域,被称为尾巴,对于RV种类A-C和G-J的自低聚是必需的。除了RVJ的NSP5,所有NSP5都与它们的同源NSP2相互作用。我们还发现,物种间VLS是在密切相关的RV物种B与G和D与F之间形成的。当NSP5RVH和RVJ的尾部被来自RVA的NSP5的尾部取代并与它们各自的NSP2共表达时,形成来自RVH和RVJ的VLS。
目的:轮状病毒(RV)分为9种,A-D和F-J,感染哺乳动物和鸟类。由于缺乏研究工具,关于RV复制的所有累积知识是基于RV种类A(RVA)。RV复制区室是球形胞质结构,称为病毒质,仅在RV物种A中鉴定出,在这项研究中,我们检查了病毒质样结构(VLSs)的形成,NSP5与NSP2共表达在RV物种A到J。B,D,F,G,而我,而RV物种C形成丝状结构。RV物种H和J不与其同源NSP5和NSP2形成VLS。类似于RVA,NSP5在所有RV物种中自我寡聚化,这是VLS形成所需的。本研究提供了非RVA复制机制的基本知识,这可能有助于制定策略来阻止跨RV物种的病毒感染。
Rotaviruses (RVs) are classified into nine species, A-D and F-J, with species A being the most studied. In rotavirus of species A (RVA), replication occurs in viroplasms, which are cytosolic globular inclusions composed of main building block proteins NSP5, NSP2, and VP2. The co-expression of NSP5 with either NSP2 or VP2 in uninfected cells leads to the formation of
viroplasm-like structures (VLSs). Although morphologically identical to viroplasms, VLSs do not produce viral progeny but serve as excellent tools for studying complex viroplasms. A knowledge gap exists regarding non-RVA viroplasms due to the lack of specific antibodies and suitable cell culture systems. In this study, we explored the ability of NSP5 and NSP2 from non-RVA species to form VLSs. The co-expression of these two proteins led to globular VLSs in RV species A, B, D, F, G, and I, while RVC formed filamentous VLSs. The co-expression of NSP5 and NSP2 of RV species H and J did not result in VLS formation. Interestingly, NSP5 of all RV species self-oligomerizes, with the ordered C-terminal region, termed the tail, being necessary for self-oligomerization of RV species A-C and G-J. Except for NSP5 from RVJ, all NSP5 interacted with their cognate NSP2. We also found that interspecies VLS are formed between closely related RV species B with G and D with F. Additionally, VLS from RVH and RVJ formed when the tail of NSP5 RVH and RVJ was replaced by the tail of NSP5 from RVA and co-expressed with their respective NSP2.
OBJECTIVE: Rotaviruses (RVs) are classified into nine species, A-D and F-J, infecting mammals and birds. Due to the lack of research tools, all cumulative knowledge on RV replication is based on RV species A (RVA). The RV replication compartments are globular cytosolic structures named viroplasms, which have only been identified in RV species A. In this study, we examined the formation of
viroplasm-like structures (VLSs) by the co-expression of NSP5 with NSP2 across RV species A to J. Globular VLSs formed for RV species A, B, D, F, G, and I, while RV species C formed filamentous structures. The RV species H and J did not form VLS with their cognates NSP5 and NSP2. Similar to RVA, NSP5 self-oligomerizes in all RV species, which is required for VLS formation. This study provides basic knowledge of the non-RVA replication mechanisms, which could help develop strategies to halt virus infection across RV species.