Molecular architecture of coronavirus double-membrane vesicle pore complex

• 成果类型: Article
• 署名作者: Huang, Yixin; Wang, Tongyun; Zhong, Lijie; Zhang, Wenxin; Zhang, Yu; Yu, Xiulian; Yuan, Shuofeng; Ni, Tao
• 署名单位: University of Hong Kong; University of Hong Kong; Hong Kong Polytechnic University; University of Hong Kong
• 刊物名称: Nature
• ISSN/ISSBN: 0028-5896
• DOI: 10.1038/s41586-024-07817-y
• 发表日期: 2024-09-05
• 页码: 224-+
• 关键词: tomography; nsp4

摘要

Coronaviruses remodel the intracellular host membranes during replication, forming double-membrane vesicles (DMVs) to accommodate viral RNA synthesis and modifications(1,2). SARS-CoV-2 non-structural protein 3 (nsp3) and nsp4 are the minimal viral components required to induce DMV formation and to form a double-membrane-spanning pore, essential for the transport of newly synthesized viral RNAs3-5. The mechanism of DMV pore complex formation remains unknown. Here we describe the molecular architecture of the SARS-CoV-2 nsp3-nsp4 pore complex, as resolved by cryogenic electron tomography and subtomogram averaging in isolated DMVs. The structures uncover an unexpected stoichiometry and topology of the nsp3-nsp4 pore complex comprising 12 copies each of nsp3 and nsp4, organized in 4 concentric stacking hexamer rings, mimicking a miniature nuclear pore complex. The transmembrane domains are interdigitated to create a high local curvature at the double-membrane junction, coupling double-membrane reorganization with pore formation. The ectodomains form extensive contacts in a pseudo-12-fold symmetry, belting the pore complex from the intermembrane space. A central positively charged ring of arginine residues coordinates the putative RNA translocation, essential for virus replication. Our work establishes a framework for understanding DMV pore formation and RNA translocation, providing a structural basis for the development of new antiviral strategies to combat coronavirus infection.