Katoh Hiroshi, Kimura Ryuichi, Sekizuka Tsuyoshi, Matsuoka Kohei, Hosogi Mika, Kitai Yuki, Akahori Yukiko, Kato Fumihiro, Kataoka Michiyo, Kobayashi Hirotaka, Nagata Noriyo, Suzuki Tadaki, Ohkawa Yasuyuki, Oki Shinya, Takeda Makoto
Department of Microbiology, Graduate School of Medicine and Faculty of Medicine, The University of Tokyo, Tokyo 113-0033, Japan.
Institute of Resource Development and Analysis, Kumamoto University, Kumamoto 860-0811, Japan.
Sci Adv. 2024 Dec 6;10(49):eadr0359. doi: 10.1126/sciadv.adr0359.
Viral RNA synthesis of mononegaviruses occurs in cytoplasmic membraneless organelles called inclusion bodies (IBs). Here, we report that IBs of mumps virus (MuV), which is the causative agent of mumps and belongs to the family , displayed liquid organelle properties formed by liquid-liquid phase separation. Super-resolution microscopic analysis of MuV IBs demonstrated that nucleocapsid and phospho (P)-proteins formed a cage-like structure and that the viral polymerase adopted a reticular pattern and colocalized with viral RNAs. In addition, we characterized host RNAs localized in MuV IBs by a spatial transcriptome analysis, and found that RNAs containing G-quadruplex motif sequences (G4-RNAs) were concentrated. An in vitro phase separation assay showed that the G4-RNAs interacted with the P protein and enhanced condensation in P droplets. Together, our data show that MuV generates IBs with a characteristic cage-like structure and host G4-RNAs play an important role in forming MuV IBs.
单股负链RNA病毒的病毒RNA合成发生在称为包涵体(IBs)的细胞质无膜细胞器中。在此,我们报告称,腮腺炎病毒(MuV)(腮腺炎的病原体,属于 科)的包涵体表现出由液-液相分离形成的液体细胞器特性。对MuV包涵体的超分辨率显微镜分析表明,核衣壳蛋白和磷(P)蛋白形成了笼状结构,并且病毒聚合酶呈现出网状模式并与病毒RNA共定位。此外,我们通过空间转录组分析对定位在MuV包涵体中的宿主RNA进行了表征,发现含有G-四链体基序序列的RNA(G4-RNAs)被富集。体外相分离试验表明,G4-RNAs与P蛋白相互作用并增强了P液滴中的凝聚。总之,我们的数据表明MuV产生具有特征性笼状结构的包涵体,并且宿主G4-RNAs在形成MuV包涵体中起重要作用。