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TitleCryo-EM Structure of an Extended SARS-CoV-2 Replication and Transcription Complex Reveals an Intermediate State in Cap Synthesis.
Journal, issue, pagesCell, Vol. 184, Issue 1, Page 184-193.e10, Year 2021
Publish dateJan 7, 2021
AuthorsLiming Yan / Ji Ge / Litao Zheng / Ying Zhang / Yan Gao / Tao Wang / Yucen Huang / Yunxiang Yang / Shan Gao / Mingyu Li / Zhenyu Liu / Haofeng Wang / Yingjian Li / Yu Chen / Luke W Guddat / Quan Wang / Zihe Rao / Zhiyong Lou /
PubMed AbstractTranscription of SARS-CoV-2 mRNA requires sequential reactions facilitated by the replication and transcription complex (RTC). Here, we present a structural snapshot of SARS-CoV-2 RTC as it ...Transcription of SARS-CoV-2 mRNA requires sequential reactions facilitated by the replication and transcription complex (RTC). Here, we present a structural snapshot of SARS-CoV-2 RTC as it transitions toward cap structure synthesis. We determine the atomic cryo-EM structure of an extended RTC assembled by nsp7-nsp8-nsp12-nsp13-RNA and a single RNA-binding protein, nsp9. Nsp9 binds tightly to nsp12 (RdRp) NiRAN, allowing nsp9 N terminus inserting into the catalytic center of nsp12 NiRAN, which then inhibits activity. We also show that nsp12 NiRAN possesses guanylyltransferase activity, catalyzing the formation of cap core structure (GpppA). The orientation of nsp13 that anchors the 5' extension of template RNA shows a remarkable conformational shift, resulting in zinc finger 3 of its ZBD inserting into a minor groove of paired template-primer RNA. These results reason an intermediate state of RTC toward mRNA synthesis, pave a way to understand the RTC architecture, and provide a target for antiviral development.
External linksCell / PubMed:33232691 / PubMed Central
MethodsEM (single particle)
Resolution2.83 Å
Structure data

30504

7cyq

EMDB-30504, PDB-7cyq:
Cryo-EM structure of an extended SARS-CoV-2 replication and transcription complex reveals an intermediate state in cap synthesis
Method: EM (single particle) / Resolution: 2.83 Å

Chemicals

ChemComp-ZN:
Unknown entry

ChemComp-GDP:
GUANOSINE-5'-DIPHOSPHATE / GDP, energy-carrying molecule*YM / Guanosine diphosphate

ChemComp-MG:
Unknown entry

Source
  • severe acute respiratory syndrome coronavirus 2
KeywordsVIRAL PROTEIN/RNA / SARS-CoV-2 / Transcription and replication / nsp9 / nsp13 / nsp12-nsp7-nsp8 / VIRAL PROTEIN-RNA complex

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