2. Academic Validation
  3. SARS-CoV-2 NSP3/4 control formation of replication organelle and recruitment of RNA polymerase NSP12

SARS-CoV-2 NSP3/4 control formation of replication organelle and recruitment of RNA polymerase NSP12

  • J Cell Biol. 2025 Mar 3;224(3):e202306101. doi: 10.1083/jcb.202306101.
Jinping Yang # 1 2,  Buyun Tian # 2,  Pei Wang # 2,  Rongrong Chen # 2,  Ke Xiao 3,  Xubing Long 2,  Xinyue Zheng 4,  Yun Zhu 3,  Fei Sun 3 4,  Yongxia Shi 5,  Yaming Jiu 4 6,  Wei Ji 3,  Yanhong Xue 3 4,  Tao Xu 2 3 4,  Zonghong Li 2
Affiliations

Affiliations

  • 1 Key Laboratory of Molecular Biophysics of the Ministry of Education, College of Life Science and Technology, Huazhong University of Science and Technology, Wuhan, China.
  • 2 Guangzhou National Laboratory , Guangzhou, China.
  • 3 National Laboratory of Biomacromolecules, CAS Center for Excellence in Biomacromolecules, Institute of Biophysics, Chinese Academy of Sciences, Beijing, China.
  • 4 College of Life Sciences, University of Chinese Academy of Sciences , Beijing, China.
  • 5 State Key Laboratory of Respiratory Disease, Health Quarantine Institute of IQTC, Guangzhou Customs District, Guangzhou, China.
  • 6 Unit of Cell Biology and Imaging Study of Pathogen Host Interaction, Key Laboratory of Molecular Virology and Immunology, Shanghai Institute of Immunity and Infection, Chinese Academy of Sciences , Shanghai, China.
  • # Contributed equally.
PMID: 39737877 DOI: 10.1083/jcb.202306101
Abstract

β-coronavirus rearranges the host cellular membranes to form double-membrane vesicles (DMVs) via NSP3/4, which anchor replication-transcription complexes (RTCs), thereby constituting the replication organelles (ROs). However, the impact of specific domains within NSP3/4 on DMV formation and RO assembly remains largely unknown. By using cryogenic-correlated light and electron microscopy (cryo-CLEM), we discovered that the N-terminal and C-terminal domains (NTD and CTD) of SARS-CoV-2 NSP3 are essential for DMV formation. Nevertheless, the CTD of NSP4 is not essential for DMV formation but regulates the DMV numbers. Additionally, the NTD of NSP3 is required for recruiting the RTC component to the cytosolic face of DMVs through direct interaction with NSP12 to assemble ROs. Furthermore, we observed that the size of NSP3/4-induced DMVs is smaller than virus-induced DMVs and established that RTC-mediated synthesis of double-stranded RNA (dsRNA) cargo plays a crucial role in determining DMV size. Collectively, our findings reveal that β-coronaviruses exploit the NSP3/4/12 axis to establish the viral ROs.

Figures
Products