Coupling of N7-methyltransferase and 3′-5′ exoribonuclease with SARS-CoV-2 polymerase reveals mechanisms for capping and proofreading

被引：121

作者：

Yan, Liming ^{[1
,2
,3
]}

Yang, Yunxiang ^{[1
,5
]}

Li, Mingyu ^{[1
]}

Zhang, Ying ^{[1
]}

Zheng, Litao ^{[2
,3
,4
]}

Ge, Ji ^{[4
]}

Huang, Yucen C. ^{[1
]}

Liu, Zhenyu ^{[1
]}

Wang, Tao ^{[1
,2
,3
]}

Gao, Shan ^{[1
]}

Zhang, Ran ^{[4
]}

Huang, Yuanyun Y. ^{[6
]}

Guddat, Luke W. ^{[7
]}

Gao, Yan ^{[2
,3
]}

Rao, Zihe ^{[1
,2
,3
,4
,8
,9
,10
,11
]}

Lou, Zhiyong ^{[1
,11
]}

机构：

[1] Tsinghua Univ, Sch Med, MOE Key Lab Prot Sci, Beijing, Peoples R China

[2] ShanghaiTech Univ, Shanghai Inst Adv Immunochem Studies, Shanghai, Peoples R China

[3] ShanghaiTech Univ, Sch Life Sci & Technol, Shanghai, Peoples R China

[4] Tsinghua Univ, Sch Life Sci, Beijing, Peoples R China

[5] Tsinghua Univ, Tsinghua Univ Peking Univ Joint Ctr Life Sci, Sch Life Sci, Beijing, Peoples R China

[6] CALTECH, Div Chem & Chem Engn, Pasadena, CA 91125 USA

[7] Univ Queensland, Sch Chem & Mol Biosci, Brisbane, Qld, Australia

[8] Nankai Univ, Coll Life Sci, State Key Lab Med Chem Biol, Tianjin, Peoples R China

[9] Nankai Univ, Coll Pharm, Tianjin, Peoples R China

[10] Chinese Acad Sci, CAS Ctr Excellence Biomacromol, Inst Biophys, Natl Lab Biomacromol, Beijing, Peoples R China

[11] Guangzhou Lab, Guangzhou, Peoples R China

来源：

CELL | 2021年 / 184卷 / 13期

基金：

中国国家自然科学基金;

关键词：

CRYO-EM STRUCTURE; STRUCTURAL BASIS; RNA-POLYMERASE; HIGH-FIDELITY; REPLICATION; TRANSCRIPTION; VIRUS; NSP14; BACKTRACKING; EXCISION;

D O I：

10.1016/j.cell.2021.05.033

中图分类号：

Q5 [生物化学]; Q7 [分子生物学];

学科分类号：

071010 ; 081704 ;

摘要：

The capping of mRNA and the proofreading play essential roles in SARS-CoV-2 replication and transcription. Here, we present the cryo-EM structure of the SARS-CoV-2 replication-transcription complex (RTC) in a form identified as Cap(0)-RTC, which couples a co-transcriptional capping complex (CCC) composed of nsp12 NiRAN, nsp9, the bifunctional nsp14 possessing an N-terminal exoribonuclease (ExoN) and a C-terminal N7-methyltransferase (N7-MTase), and nsp10 as a cofactor of nsp14. Nsp9 and nsp12 NiRAN recruit nsp10/nsp14 into the Cap(0)-RTC, forming the N7-CCC to yield cap(0) ((7Me)GpppA) at 5' end of pre-mRNA. A dimeric form of Cap(0)-RTC observed by cryo-EM suggests an in trans backtracking mechanism for nsp14 ExoN to facilitate proofreading of the RNA in concert with polymerase nsp12. These results not only provide a structural basis for understanding co-transcriptional modification of SARS-CoV-2 mRNA but also shed light on how replication fidelity in SARS-CoV-2 is maintained.

引用

页码：3474 / +

页数：23

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