The nucleotide addition cycle of the SARS-CoV-2 polymerase.
Journal
bioRxiv : the preprint server for biology
Titre abrégé: bioRxiv
Pays: United States
ID NLM: 101680187
Informations de publication
Date de publication:
27 Mar 2021
27 Mar 2021
Historique:
pubmed:
2
4
2021
medline:
2
4
2021
entrez:
1
4
2021
Statut:
epublish
Résumé
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 have used 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 the first evidence that an RdRp uses a thermal ratchet instead of a power stroke to transition from the pre- to post-translocated state. Ultra-stable magnetic tweezers enables the direct observation of coronavirus polymerase deep and long-lived backtrack that are strongly stimulated by secondary structure in the template. The framework presented 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.
Identifiants
pubmed: 33791706
doi: 10.1101/2021.03.27.437309
pmc: PMC8010733
pii:
doi:
Types de publication
Preprint
Langues
eng
Subventions
Organisme : NCI NIH HHS
ID : P30 CA016086
Pays : United States
Organisme : NIAID NIH HHS
ID : R01 AI158463
Pays : United States
Commentaires et corrections
Type : UpdateIn
Déclaration de conflit d'intérêts
Declaration of interests: The authors declare no competing interest
Références
Annu Rev Virol. 2015 Nov;2(1):265-88
pubmed: 26958916
Structure. 2012 Sep 5;20(9):1519-27
pubmed: 22819218
Nature. 2020 Aug;584(7819):154-156
pubmed: 32438371
Nat Commun. 2018 Jul 26;9(1):2930
pubmed: 30050038
Nat Commun. 2020 Nov 18;11(1):5874
pubmed: 33208736
J Virol. 2007 Nov;81(22):12135-44
pubmed: 17804504
Nat Commun. 2019 May 28;10(1):2342
pubmed: 31138817
Cell. 2021 Jan 7;184(1):184-193.e10
pubmed: 33232691
Proc Natl Acad Sci U S A. 2010 Dec 28;107(52):22505-10
pubmed: 21148772
PLoS Pathog. 2013 Aug;9(8):e1003565
pubmed: 23966862
Nucleic Acids Res. 2012 Jul;40(13):6174-86
pubmed: 22434889
Biophys J. 2009 Mar 18;96(6):2189-93
pubmed: 19289045
Nucleic Acids Res. 2012 Jul;40(13):6187-98
pubmed: 22434886
N Engl J Med. 2021 Feb 11;384(6):497-511
pubmed: 33264556
Nat Commun. 2020 Sep 17;11(1):4682
pubmed: 32943628
Nature. 2000 Mar 2;404(6773):103-6
pubmed: 10716452
Nucleic Acids Res. 2020 Jun 4;48(10):5591-5602
pubmed: 32286652
Proc Natl Acad Sci U S A. 2021 May 11;118(19):
pubmed: 33883267
iScience. 2020 Dec 18;23(12):101849
pubmed: 33283177
J Mol Biol. 2014 Apr 3;426(7):1407-19
pubmed: 24424421
J Am Chem Soc. 2007 Dec 5;129(48):14966-73
pubmed: 17997555
Biophys J. 2015 Nov 17;109(10):2113-25
pubmed: 26588570
PLoS Pathog. 2010 May 06;6(5):e1000896
pubmed: 20463816
Proc Natl Acad Sci U S A. 2018 Jan 9;115(2):E162-E171
pubmed: 29279395
Mol Cell. 2020 Sep 3;79(5):710-727
pubmed: 32853546
Nat Commun. 2020 May 25;11(1):2605
pubmed: 32451382
Proc Natl Acad Sci U S A. 2012 May 22;109(21):8115-20
pubmed: 22573817
Commun Biol. 2021 Aug 24;4(1):999
pubmed: 34429502
Adv Virus Res. 2016;96:59-126
pubmed: 27712628
Biophys J. 2017 Feb 28;112(4):575-583
pubmed: 28256218
Annu Rev Biochem. 2004;73:705-48
pubmed: 15189157
J Biol Chem. 2014 Oct 24;289(43):29531-44
pubmed: 25213864
Cell Rep. 2017 Oct 24;21(4):1063-1076
pubmed: 29069588
Cell Rep. 2015 Feb 17;10(6):983-992
pubmed: 25683720
Nat Rev Microbiol. 2013 May;11(5):327-36
pubmed: 23524517
Nucleic Acids Res. 2015 Dec 2;43(21):10421-9
pubmed: 26496948
Data Brief. 2020 Mar 12;30:105397
pubmed: 32258273
Science. 2020 May 15;368(6492):779-782
pubmed: 32277040
Biochemistry. 2004 May 11;43(18):5126-37
pubmed: 15122878
Nat Med. 2000 Dec;6(12):1375-9
pubmed: 11100123
EMBO J. 2009 Sep 16;28(18):2794-802
pubmed: 19661923
Nature. 2005 Nov 24;438(7067):460-5
pubmed: 16284617
Biophys J. 2019 Aug 6;117(3):509-519
pubmed: 31337545
Elife. 2013 Sep 24;2:e00971
pubmed: 24066225
Cell. 2020 Jul 23;182(2):417-428.e13
pubmed: 32526208
Sci Rep. 2018 Oct 29;8(1):15920
pubmed: 30374099
Nature. 2003 Dec 11;426(6967):684-7
pubmed: 14634670
Proc Natl Acad Sci U S A. 2014 Oct 28;111(43):15408-13
pubmed: 25313077
Nat Commun. 2015 Jan 07;6:5885
pubmed: 25565216
Cell. 2020 Sep 17;182(6):1560-1573.e13
pubmed: 32783916