Details

Autor(en) / Beteiligte

• Wang, Quan
• Wu, Jiqin
• Wang, Haofeng
• Gao, Yan
• Liu, Qiaojie
• Mu, An
• Ji, Wenxin
• Yan, Liming
• Zhu, Yan
• Zhu, Chen
• Fang, Xiang
• Yang, Xiaobao
• Huang, Yucen
• Gao, Hailong
• Liu, Fengjiang
• Ge, Ji
• Sun, Qianqian
• Yang, Xiuna
• Xu, Wenqing
• Liu, Zhijie
• Yang, Haitao
• Lou, Zhiyong
• Jiang, Biao
• Guddat, Luke W.
• Gong, Peng
• Rao, Zihe

Titel

Structural Basis for RNA Replication by the SARS-CoV-2 Polymerase

Ist Teil von

• Cell, 2020-07, Vol.182 (2), p.417-428.e13

Ort / Verlag

United States: Elsevier Inc

Erscheinungsjahr

2020

Link zum Volltext

Quelle

EZB Electronic Journals Library

Beschreibungen/Notizen

• Nucleotide analog inhibitors, including broad-spectrum remdesivir and favipiravir, have shown promise in in vitro assays and some clinical studies for COVID-19 treatment, this despite an incomplete mechanistic understanding of the viral RNA-dependent RNA polymerase nsp12 drug interactions. Here, we examine the molecular basis of SARS-CoV-2 RNA replication by determining the cryo-EM structures of the stalled pre- and post- translocated polymerase complexes. Compared with the apo complex, the structures show notable structural rearrangements happening to nsp12 and its co-factors nsp7 and nsp8 to accommodate the nucleic acid, whereas there are highly conserved residues in nsp12, positioning the template and primer for an in-line attack on the incoming nucleotide. Furthermore, we investigate the inhibition mechanism of the triphosphate metabolite of remdesivir through structural and kinetic analyses. A transition model from the nsp7-nsp8 hexadecameric primase complex to the nsp12-nsp7-nsp8 polymerase complex is also proposed to provide clues for the understanding of the coronavirus transcription and replication machinery. [Display omitted] •Structures of SARS-CoV-2 RNA polymerase in complexes with RNA revealed•Conformational changes in nsp8 and its interaction with the exiting RNA are observed•Incorporation and delayed-chain-termination mechanism of remdesivir is elucidated•Transition model from primase complex to polymerase complex is proposed Cryo-EM structures of the SARS-CoV-2 RNA polymerase in complexes with RNA, before and after RNA translocation, reveals structural rearrangements that the RNA-dependent RNA polymerase (RdRp) nsp12 and its co-factors (nsp7 and nsp8) undergo to accommodate nucleic acid binding. Further insights into how the complex is inhibited by remdesivir, and into the primase to polymerase transition, are also presented.