The nucleotide addition cycle of the SARS-CoV-2 polymerase

被引：18

作者：

Bera, Subhas Chandra ^{[1
]}

Seifert, Mona ^{[1
]}

Kirchdoerfer, Robert N. ^{[2
,3
]}

van Nies, Pauline ^{[1
]}

Wubulikasimu, Yibulayin ^{[1
]}

Quack, Salina ^{[1
]}

Papini, Flavia S. ^{[1
]}

Arnold, Jamie J. ^{[6
]}

Canard, Bruno ^{[4
,5
]}

Cameron, Craig E. ^{[6
]}

Depken, Martin ^{[7
]}

Dulin, David ^{[1
,8
,9
]}

机构：

[1] Friedrich Alexander Univ Erlangen Nurnberg FAU, Interdisciplinary Ctr Clin Res, Jr Res Grp 2, Cauerstr 3, D-91058 Erlangen, Germany

[2] Univ Wisconsin, Dept Biochem, Madison, WI 53706 USA

[3] Univ Wisconsin, Inst Mol Virol, Madison, WI 53706 USA

[4] CNRS, Architecture & Fonct Macromol Biol, Polytech Case 925, F-13009 Marseille, France

[5] Aix Marseille Univ, UMR 7257, Polytech Case 925, F-13009 Marseille, France

[6] Univ N Carolina, Sch Med, Dept Microbiol & Immunol, Chapel Hill, NC 27599 USA

[7] Delft Univ Technol, Kavli Inst Nanosci, Dept Bionanosci, Van Maasweg 9, NL-2629 HZ Delft, Netherlands

[8] Vrije Univ Amsterdam, Dept Phys & Astron, De Boelelaan 1081, NL-1081 NV Amsterdam, Netherlands

[9] Vrije Univ Amsterdam, LaserLaB Amsterdam, De Boelelaan 1081, NL-1081 NV Amsterdam, Netherlands

来源：

CELL REPORTS | 2021年 / 36卷 / 09期

基金：

欧盟地平线“2020”;

关键词：

STRUCTURAL BASIS; SINGLE-MOLECULE; RNA-SYNTHESIS; REPLICATION; FIDELITY; TRANSLOCATION; BACKTRACKING; MECHANISM; DYNAMICS; HELICASE;

D O I：

10.1016/j.celrep.2021.109650

中图分类号：

Q2 [细胞生物学];

学科分类号：

071009 ; 090102 ;

摘要：

Coronaviruses have evolved elaborate multisubunit machines to replicate and transcribe their genomes. Central to these machines are the RNA-dependent RNA polymerase subunit (nsp12) and its intimately associated cofactors (nsp7 and nsp8). We use a high-throughput magnetic-tweezers approach to develop a mechanochemical description of this core polymerase. The core polymerase exists in at least three catalytically distinct conformations, one being kinetically consistent with incorporation of incorrect nucleotides. We provide evidence that the RNA-dependent RNA polymerase (RdRp) uses a thermal ratchet instead of a power stroke to transition from the pre- to post-translocated state. Ultra-stable magnetic tweezers enable the direct observation of coronavirus polymerase deep and long-lived backtracking that is strongly stimulated by secondary structures in the template. The framework we present here elucidates one of the most important structure-dynamics-function relationships in human health today and will form the grounds for understanding the regulation of this complex.

引用

页数：22

共 63 条

[31] Structure of the SARS-CoV nsp12 polymerase bound to nsp7 and nsp8 co-factors [J].