SARS-CoV-2 NSP3/4 control formation of replication organelle and recruitment of RNA polymerase NSP12
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.
Authors
- Yongxia Shi (ORCID: https://orcid.org/0000-0003-3590-2365)
- Ke Xiao
- Tao Xu (ORCID: https://orcid.org/0000-0002-8260-9754)
- Yaming Jiu (ORCID: https://orcid.org/0000-0002-8601-8820)
- Yanhong Xue (ORCID: https://orcid.org/0000-0003-3214-4175)
- Zonghong Li (ORCID: https://orcid.org/0000-0002-5153-0007)
- Buyun Tian (ORCID: https://orcid.org/0000-0002-0027-5499)
- Yun Zhu (ORCID: https://orcid.org/0000-0001-9382-8592)
- Xinyue Zheng
- Fei Sun (ORCID: https://orcid.org/0000-0002-0351-5144)
- Pei Wang (ORCID: https://orcid.org/0000-0001-5612-705X)
- Rongrong Chen (ORCID: https://orcid.org/0009-0001-9646-0938)
- Jinping Yang
- Wei Ji (ORCID: https://orcid.org/0000-0002-2317-9053)
- Xueqing Long
Institutions
- Chinese Academy of Sciences (CN)
- Guangzhou Experimental Station (CN)
- Institute of Biophysics (CN)
- State Key Laboratory of Respiratory Disease (CN)
- Guangdong Inspection and Quarantine Technology Center (China) (CN)
- University of Chinese Academy of Sciences (CN)
- Huazhong University of Science and Technology (CN)
Publication Details
- Journal
- The Journal of Cell Biology
- Published
- 2024-12-31
- DOI
- https://doi.org/10.1083/jcb.202306101
- Citations
- 18
- Primary Topic
- SARS-CoV-2 and COVID-19 Research
- Type
- article
- Field-Weighted Citation Impact
- 7.00
Funders
- National Natural Science Foundation of China
- Chinese Academy of Sciences
- National Key Research and Development Program of China