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

Liming Yan; Yunxiang Yang; Mingyu Li; Ying Zhang; Litao Zheng; Ji Ge; Yucen C. Huang; Zhenyu Liu; Tao Wang; Shan Gao; Ran Zhang; Yuanyun Y. Huang; Luke W. Guddat; Yan Gao; Zihe Rao; Zhiyong Lou

doi:10.1016/j.cell.2021.05.033

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

Liming Yan, Yunxiang Yang, Mingyu Li, Ying Zhang, Litao Zheng, Ji Ge, Yucen C. Huang, Zhenyu Liu, Tao Wang, Shan Gao, Ran Zhang, Yuanyun Y. Huang, Luke W. Guddat, Yan Gao, Zihe Rao, Zhiyong Lou

Résultat de recherche: Article › examen par les pairs

128 Citations (Scopus)

Résumé

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) (^7MeGpppA) 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.

Langue d'origine	English
Pages (de-à)	3474-3485.e11
Journal	Cell
Volume	184
Numéro de publication	13
DOI	https://doi.org/10.1016/j.cell.2021.05.033
Statut de publication	Published - juin 24 2021
Publié à l'externe	Oui

Note bibliographique

Funding Information:
This work was supported by the National Natural Science Foundation of China ( U20A20135 ), the National Program on Key Research Project of China ( 2020YFA0707500 and 2017YFC0840300 ), Tsinghua University Spring Breeze Fund ( 2020Z99CFG015 ), and the Strategic Priority Research Program of the Chinese Academy of Sciences ( XDB37030201 ).

Publisher Copyright:
© 2021 Elsevier Inc.

ASJC Scopus Subject Areas

General Biochemistry,Genetics and Molecular Biology

PubMed: MeSH publication types

Journal Article
Research Support, Non-U.S. Gov't

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10.1016/j.cell.2021.05.033

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Yan, L., Yang, Y., Li, M., Zhang, Y., Zheng, L., Ge, J., Huang, Y. C., Liu, Z., Wang, T., Gao, S., Zhang, R., Huang, Y. Y., Guddat, L. W., Gao, Y., Rao, Z., & Lou, Z. (2021). Coupling of N7-methyltransferase and 3′-5′ exoribonuclease with SARS-CoV-2 polymerase reveals mechanisms for capping and proofreading. Cell, 184(13), 3474-3485.e11. https://doi.org/10.1016/j.cell.2021.05.033

Yan, L, Yang, Y, Li, M, Zhang, Y, Zheng, L, Ge, J, Huang, YC, Liu, Z, Wang, T, Gao, S, Zhang, R, Huang, YY, Guddat, LW, Gao, Y, Rao, Z & Lou, Z 2021, 'Coupling of N7-methyltransferase and 3′-5′ exoribonuclease with SARS-CoV-2 polymerase reveals mechanisms for capping and proofreading', Cell, vol. 184, n° 13, pp. 3474-3485.e11. https://doi.org/10.1016/j.cell.2021.05.033

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

abstract = "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) (7MeGpppA) 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.",

author = "Liming Yan and Yunxiang Yang and Mingyu Li and Ying Zhang and Litao Zheng and Ji Ge and Huang, {Yucen C.} and Zhenyu Liu and Tao Wang and Shan Gao and Ran Zhang and Huang, {Yuanyun Y.} and Guddat, {Luke W.} and Yan Gao and Zihe Rao and Zhiyong Lou",

note = "Funding Information: This work was supported by the National Natural Science Foundation of China ( U20A20135 ), the National Program on Key Research Project of China ( 2020YFA0707500 and 2017YFC0840300 ), Tsinghua University Spring Breeze Fund ( 2020Z99CFG015 ), and the Strategic Priority Research Program of the Chinese Academy of Sciences ( XDB37030201 ). Publisher Copyright: {\textcopyright} 2021 Elsevier Inc.",

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doi = "10.1016/j.cell.2021.05.033",

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T1 - Coupling of N7-methyltransferase and 3′-5′ exoribonuclease with SARS-CoV-2 polymerase reveals mechanisms for capping and proofreading

AU - Yan, Liming

AU - Yang, Yunxiang

AU - Li, Mingyu

AU - Zhang, Ying

AU - Zheng, Litao

AU - Ge, Ji

AU - Huang, Yucen C.

AU - Liu, Zhenyu

AU - Wang, Tao

AU - Gao, Shan

AU - Zhang, Ran

AU - Huang, Yuanyun Y.

AU - Guddat, Luke W.

AU - Gao, Yan

AU - Rao, Zihe

AU - Lou, Zhiyong

N1 - Funding Information: This work was supported by the National Natural Science Foundation of China ( U20A20135 ), the National Program on Key Research Project of China ( 2020YFA0707500 and 2017YFC0840300 ), Tsinghua University Spring Breeze Fund ( 2020Z99CFG015 ), and the Strategic Priority Research Program of the Chinese Academy of Sciences ( XDB37030201 ). Publisher Copyright: © 2021 Elsevier Inc.

PY - 2021/6/24

Y1 - 2021/6/24

N2 - 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) (7MeGpppA) 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.

AB - 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) (7MeGpppA) 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.

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JO - Cell

JF - Cell

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ER -

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

Résumé

Note bibliographique

ASJC Scopus Subject Areas

PubMed: MeSH publication types

Accès au document

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