Ultrastructure of influenza virus ribonucleoprotein complexes during viral RNA synthesis.
Ultrastructure of influenza virus ribonucleoprotein complexes during viral RNA synthesis.
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DOI：
10.1038/s42003-021-02388-4
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发表时间：
2021-07-09
影响因子：
5.9
通讯作者：
Noda T
中科院分区：
文献类型：
--
作者：
Nakano M;Sugita Y;Kodera N;Miyamoto S;Muramoto Y;Wolf M;Noda T
甲型流感病毒的单链负义病毒基因组RNA（vRNA）被病毒核蛋白（NP）和RNA聚合酶包裹，形成具有螺旋杆状结构的核糖核蛋白复合物（vRNP）。vRNP负责vRNA的转录和复制。然而，RNA合成过程中的vRNP构象还没有很好地理解。在这里，使用高速原子力显微镜和冷冻电子显微镜，我们研究了体外RNA合成过程中甲型流感病毒vRNP的天然结构。观察到两种不同类型的vRNP与新合成的RNA相关联：与折叠RNA连接的完整的螺旋杆状vRNP和与环状RNA相关的变形vRNP。有趣的是，环状RNA是双链RNA，其可能包含新生RNA和从vRNP的NP分离的模板RNA。这些结果表明，虽然一些vRNP在RNA合成过程中保持其螺旋结构，但对于RNA合成的重复循环，其他vRNP意外地变得结构变形，这可能导致无法开始或继续RNA合成。因此，我们的研究结果提供了RNA合成过程中vRNP的超微结构特征。中野等人使用高速原子力显微镜和冷冻电镜研究流感病毒RNA的体外合成。他们观察到两种类型的RNA的合成：与完整的病毒核糖核蛋白（vRNP）相关的折叠单链RNA和与部分重排的vRNP结构相关的环状双链RNA。
The single-stranded, negative-sense, viral genomic RNA (vRNA) of influenza A virus is encapsidated by viral nucleoproteins (NPs) and an RNA polymerase to form a ribonucleoprotein complex (vRNP) with a helical, rod-shaped structure. The vRNP is responsible for transcription and replication of the vRNA. However, the vRNP conformation during RNA synthesis is not well understood. Here, using high-speed atomic force microscopy and cryo-electron microscopy, we investigated the native structure of influenza A vRNPs during RNA synthesis in vitro. Two distinct types of vRNPs were observed in association with newly synthesized RNAs: an intact, helical rod-shaped vRNP connected with a folded RNA and a deformed vRNP associated with a looped RNA. Interestingly, the looped RNA was a double-stranded RNA, which likely comprises a nascent RNA and the template RNA detached from NPs of the vRNP. These results suggest that while some vRNPs keep their helical structures during RNA synthesis, for the repeated cycle of RNA synthesis, others accidentally become structurally deformed, which likely results in failure to commence or continue RNA synthesis. Thus, our findings provide the ultrastructural feature of vRNPs during RNA synthesis. Nakano et al. use high-speed AFM and cryo-EM to study influenza virus RNA synthesis in vitro. They observe the synthesis of two types of RNA: folded single stranded RNA associated to an intact viral ribonucleoprotein (vRNP) and looped, double stranded RNA associated with a partially rearranged vRNP structure.
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