Zhang Yuanyuan, Huang Linlong, Ren Chaoqi, Wang Weiyang, Wang Xinlu, Gao Guangxia
Key Laboratory of Biomacromolecules (CAS), CAS Center for Excellence in Biomacromolecules, Institute of Biophysics, Chinese Academy of Sciences, Beijing, China.
University of Chinese Academy of Sciences, Beijing, China.
mBio. 2025 Jun 11;16(6):e0097925. doi: 10.1128/mbio.00979-25. Epub 2025 May 23.
β-Coronaviruses assemble at the endoplasmic reticulum-to-Golgi compartment and exit cells through the lysosomal trafficking pathway. However, the molecular mechanisms of virion assembly and egress are largely unknown. Here, we report that β-coronaviruses recruit endosomal sorting complexes required for transport (ESCRTs) to facilitate virion assembly and egress. The viral proteins N and M interacted with ESCRT components TSG101 and VPS28, respectively. Electron microscopy analysis revealed that virion assembly was disrupted by TSG101 knockdown at an early stage and by VPS28 knockdown at a late stage. Knockdown of ESCRT components MVB12A and CHMP6 did not affect virion assembly but inhibited virion egress. Downregulation of these ESCRT factors or VPS4A, or treatment of cells with the TSG101 antagonist tenatoprazole significantly inhibited the production of the virion-like particles of β-coronaviruses and the replication of human coronavirus OC43. These findings indicate that β-coronaviruses exploit ESCRT for virion assembly and egress and suggest that the interaction interface between ESCRT and the viruses could be a target for the development of broad-spectrum anti-coronavirus therapeutics.IMPORTANCEβ-Coronaviruses have caused disastrous pandemics and may cause serious pandemics in the future. Virion assembly and egress are critical steps in the life cycle of coronaviruses. However, despite extensive studies in the past few years, the molecular mechanisms for virion assembly and egress are still largely unknown. Here we show that β-coronaviruses recruit ESCRT components TSG101 and VPS28 for virion assembly and that ESCRT components MVB12A and CHMP6 are required for virion egress. Treatment of cells with the TSG101 antagonist inhibited the assembly of multiple β-coronaviruses. These findings indicate that ESCRT participates in β-coronavirus assembly and egress and might be a potential target for the development of broad-spectrum anti-coronavirus therapeutics.
β冠状病毒在内质网到高尔基体区室组装，并通过溶酶体运输途径离开细胞。然而，病毒粒子组装和释放的分子机制在很大程度上尚不清楚。在此，我们报告β冠状病毒招募运输所需的内体分选复合体（ESCRT）以促进病毒粒子的组装和释放。病毒蛋白N和M分别与ESCRT组分TSG101和VPS28相互作用。电子显微镜分析显示，病毒粒子组装在早期被TSG101敲低破坏，在后期被VPS28敲低破坏。ESCRT组分MVB12A和CHMP6的敲低不影响病毒粒子组装，但抑制病毒粒子释放。这些ESCRT因子或VPS4A的下调，或用TSG101拮抗剂替那拉唑处理细胞，显著抑制β冠状病毒的病毒样颗粒的产生和人冠状病毒OC43的复制。这些发现表明，β冠状病毒利用ESCRT进行病毒粒子的组装和释放，并表明ESCRT与病毒之间的相互作用界面可能是开发广谱抗冠状病毒疗法的靶点。
重要性
β冠状病毒已引发灾难性大流行，未来可能导致严重大流行。病毒粒子组装和释放是冠状病毒生命周期中的关键步骤。然而，尽管在过去几年进行了广泛研究，但病毒粒子组装和释放的分子机制在很大程度上仍不清楚。在这里我们表明，β冠状病毒招募ESCRT组分TSG101进行病毒粒子组装，并且ESCRT组分MVB12A和CHMP6是病毒粒子释放所必需的。用TSG101拮抗剂处理细胞抑制了多种β冠状病毒的组装。这些发现表明ESCRT参与β冠状病毒的组装和释放，可能是开发广谱抗冠状病毒疗法的潜在靶点。
