Kniert Justine, Terino Dante, Eaton Heather E, Lin Qi Feng, Wu Shiau-Yin, Strickfaden Hilmar, Shmulevitz Maya
Department of Medical Microbiology and Immunology, Li Ka Shing Institute of Virology, University of Alberta, Edmonton, Alberta, Canada.
Cell Imaging Core, Faculty of Medicine and Dentistry, University of Alberta, Edmonton, Alberta, Canada.
PLoS Pathog. 2025 Sep 2;21(9):e1013238. doi: 10.1371/journal.ppat.1013238. eCollection 2025 Sep.
Reoviruses coordinate their replication and assembly through intricate spatial and temporal compartmentalization within host cells. In this study, we elucidate the dynamics of mammalian orthoreovirus (reovirus) core replication and viral particle assembly. Using high-resolution immunofluorescence confocal microscopy, we tracked input cores and de novo cores, revealing that input cores initially form peripheral, OC-negative factories that migrate inward while seeding independent peripheral factories. Over time, these input factories transition into intermediate core-plus-outercapsid (OC) factories, which are essential for full virion assembly in the perinuclear region. Notably, de novo core proteins predominantly form independent peripheral factories that can merge or mix with others, resulting in interconnected networks. We further demonstrate that microtubules are dispensable for early core movement and factory formation but are crucial for the transition of mature, assembled virions into perinuclear deposits and for timely virion production. Disruption of microtubules delays full virus assembly, reducing progeny yield. Our findings reveal a complex, regulated interplay between spatial organization and cytoskeletal components during reovirus infection, providing insights into mechanisms that could be targeted for antiviral interventions.
呼肠孤病毒通过宿主细胞内复杂的空间和时间区室化来协调其复制和组装。在本研究中，我们阐明了哺乳动物正呼肠孤病毒（呼肠孤病毒）核心复制和病毒粒子组装的动态过程。利用高分辨率免疫荧光共聚焦显微镜，我们追踪了输入核心和新生核心，发现输入核心最初形成外周的、OC阴性工厂，这些工厂向内迁移，同时播种独立的外周工厂。随着时间的推移，这些输入工厂转变为中间核心加外 capsid（OC）工厂，这对于在核周区域完全组装病毒体至关重要。值得注意的是，新生核心蛋白主要形成独立的外周工厂，这些工厂可以与其他工厂合并或混合，形成相互连接的网络。我们进一步证明，微管对于早期核心运动和工厂形成是可有可无的，但对于成熟组装的病毒体向核周沉积物的转变以及及时产生病毒体至关重要。破坏微管会延迟完整病毒的组装，降低子代产量。我们的研究结果揭示了呼肠孤病毒感染期间空间组织和细胞骨架成分之间复杂的、受调控的相互作用，为抗病毒干预的潜在靶点机制提供了见解。
