Structural basis for helicase-polymerase coupling in the SARS-CoV-2 replication-transcription complex.
Journal
bioRxiv : the preprint server for biology
Titre abrégé: bioRxiv
Pays: United States
ID NLM: 101680187
Informations de publication
Date de publication:
13 Jul 2020
13 Jul 2020
Historique:
entrez:
18
7
2020
pubmed:
18
7
2020
medline:
18
7
2020
Statut:
epublish
Résumé
SARS-CoV-2 is the causative agent of the 2019-2020 pandemic. The SARS-CoV-2 genome is replicated-transcribed by the RNA-dependent RNA polymerase holoenzyme (subunits nsp7/nsp82/nsp12) along with a cast of accessory factors. One of these factors is the nsp13 helicase. Both the holo-RdRp and nsp13 are essential for viral replication and are targets for treating the disease COVID-19. Here we present cryo-electron microscopic structures of the SARS-CoV-2 holo-RdRp with an RNA template-product in complex with two molecules of the nsp13 helicase. The Nidovirus-order-specific N-terminal domains of each nsp13 interact with the N-terminal extension of each copy of nsp8. One nsp13 also contacts the nsp12-thumb. The structure places the nucleic acid-binding ATPase domains of the helicase directly in front of the replicating-transcribing holo-RdRp, constraining models for nsp13 function. We also observe ADP-Mg2+ bound in the nsp12 N-terminal nidovirus RdRp-associated nucleotidyltransferase domain, detailing a new pocket for anti-viral therapeutic development.
Identifiants
pubmed: 32676607
doi: 10.1101/2020.07.08.194084
pmc: PMC7359531
pii:
doi:
Types de publication
Preprint
Langues
eng
Subventions
Organisme : NIGMS NIH HHS
ID : P41 GM103310
Pays : United States
Organisme : NIGMS NIH HHS
ID : U24 GM129539
Pays : United States
Organisme : NIGMS NIH HHS
ID : P41 GM109824
Pays : United States
Organisme : NIGMS NIH HHS
ID : R35 GM130234
Pays : United States
Organisme : NIGMS NIH HHS
ID : P41 GM103314
Pays : United States
Organisme : NIGMS NIH HHS
ID : R35 GM118130
Pays : United States
Commentaires et corrections
Type : UpdateIn
Références
PLoS Pathog. 2017 Jun 26;13(6):e1006474
pubmed: 28651017
Nucleic Acids Res. 2015 Dec 2;43(21):10421-9
pubmed: 26496948
Cell. 1999 Sep 17;98(6):811-24
pubmed: 10499798
PLoS One. 2007 May 23;2(5):e459
pubmed: 17520018
Nature. 2020 Mar;579(7798):265-269
pubmed: 32015508
Annu Rev Virol. 2015 Nov;2(1):265-88
pubmed: 26958916
Nature. 2020 Mar;579(7798):270-273
pubmed: 32015507
Nat Methods. 2017 Aug;14(8):793-796
pubmed: 28671674
J Biol Chem. 1997 Jun 13;272(24):15329-38
pubmed: 9182561
J Virol. 2004 Jan;78(2):980-94
pubmed: 14694129
Nucleic Acids Res. 2016 Jul 8;44(W1):W351-5
pubmed: 27131377
J Virol. 2004 Jun;78(11):5619-32
pubmed: 15140959
Cell. 2012 Jun 22;149(7):1438-45
pubmed: 22726433
J Mol Biol. 2010 Jan 29;395(4):686-704
pubmed: 19895816
PLoS Pathog. 2010 May 06;6(5):e1000896
pubmed: 20463816
Mol Cell. 2019 Jul 25;75(2):298-309.e4
pubmed: 31103420
J Comput Chem. 2004 Oct;25(13):1605-12
pubmed: 15264254
Cell. 1998 May 15;93(4):627-37
pubmed: 9604937
Nat Methods. 2017 Mar;14(3):290-296
pubmed: 28165473
Trends Biochem Sci. 1990 Jan;15(1):6-10
pubmed: 2180149
Acta Crystallogr D Biol Crystallogr. 2010 Jan;66(Pt 1):12-21
pubmed: 20057044
Annu Rev Virol. 2014 Nov;1(1):111-32
pubmed: 26958717
J Virol. 2004 Jul;78(14):7833-8
pubmed: 15220459
Adv Exp Med Biol. 1998;440:215-9
pubmed: 9782283
RNA Biol. 2011 Mar-Apr;8(2):270-9
pubmed: 21593585
Nature. 2020 Aug;584(7819):154-156
pubmed: 32438371
Elife. 2017 Mar 20;6:
pubmed: 28318486
Annu Rev Microbiol. 2006;60:211-30
pubmed: 16712436
Mol Cell. 2002 Sep;10(3):647-57
pubmed: 12408831
RNA. 2000 Jul;6(7):1056-68
pubmed: 10917600
Science. 2009 May 29;324(5931):1203-6
pubmed: 19478184
Cell. 2003 Aug 8;114(3):347-57
pubmed: 12914699
J Virol. 2007 Nov;81(22):12135-44
pubmed: 17804504
Proc Natl Acad Sci U S A. 2006 Mar 28;103(13):5108-13
pubmed: 16549795
Proc Natl Acad Sci U S A. 1997 Mar 4;94(5):1755-60
pubmed: 9050851
Nucleic Acids Res. 2015 Sep 30;43(17):8416-34
pubmed: 26304538
mBio. 2011 Jun 14;2(3):e00077-11
pubmed: 21673192
Nat Commun. 2019 May 28;10(1):2342
pubmed: 31138817
Anal Chem. 2015 Apr 21;87(8):4370-6
pubmed: 25799115
Cell. 2018 Oct 18;175(3):809-821.e19
pubmed: 30270044
mBio. 2018 Mar 6;9(2):
pubmed: 29511076
Cell. 2004 Nov 12;119(4):481-9
pubmed: 15537538
Virus Res. 2006 Apr;117(1):17-37
pubmed: 16503362
Adv Virus Res. 2016;96:59-126
pubmed: 27712628
Nucleic Acids Res. 2019 Jul 9;47(12):6538-6550
pubmed: 31131400
J Virol. 2000 Jun;74(11):5213-23
pubmed: 10799597
Nature. 2011 Mar 10;471(7337):249-53
pubmed: 21346759
Cell. 1997 Apr 4;89(1):33-41
pubmed: 9094712
J Am Soc Mass Spectrom. 2019 Jan;30(1):118-127
pubmed: 29667162
Nucleic Acids Res. 2010 Nov;38(21):7626-36
pubmed: 20671029
Virus Res. 2015 Apr 16;202:12-32
pubmed: 25497126
Cell. 1993 Feb 12;72(3):459-66
pubmed: 8431948
Cell. 2009 May 29;137(5):849-59
pubmed: 19490894
Cell Rep. 2017 Oct 24;21(4):1063-1076
pubmed: 29069588
J Biol Chem. 2003 Oct 10;278(41):39578-82
pubmed: 12917423
Acta Crystallogr D Biol Crystallogr. 2004 Dec;60(Pt 12 Pt 1):2126-32
pubmed: 15572765
Science. 1993 Nov 5;262(5135):867-73
pubmed: 8235608
Acta Crystallogr D Biol Crystallogr. 2010 Feb;66(Pt 2):213-21
pubmed: 20124702
Antiviral Res. 2013 Oct;100(1):286-95
pubmed: 24012996
Elife. 2018 Nov 09;7:
pubmed: 30412051
Nucleic Acids Res. 1990 Oct 25;18(20):6097-100
pubmed: 2172928
J Virol. 2002 Feb;76(3):1293-308
pubmed: 11773405
Methods Mol Biol. 2020;2062:357-382
pubmed: 31768985
PLoS One. 2012;7(2):e32138
pubmed: 22359664
J Virol. 2005 Jan;79(2):696-704
pubmed: 15613297
Science. 2020 Jun 26;368(6498):1499-1504
pubmed: 32358203
Nucleic Acids Res. 2014 Mar;42(5):3464-77
pubmed: 24369429
J Virol. 2019 May 29;93(12):
pubmed: 30918070
J Virol. 1996 Oct;70(10):6625-33
pubmed: 8794297
J Struct Biol. 2005 Jul;151(1):41-60
pubmed: 15890530
Cell. 2003 Aug 8;114(3):335-45
pubmed: 12914698
Nat Methods. 2017 Apr;14(4):331-332
pubmed: 28250466
Elife. 2013 Sep 10;2:e01456
pubmed: 24040512
Cell. 2020 May 28;181(5):1036-1045.e9
pubmed: 32416070
Biochim Biophys Acta. 2002 Sep 13;1577(2):287-307
pubmed: 12213659
PLoS One. 2012;7(5):e36521
pubmed: 22615777
EMBO J. 2001 Dec 17;20(24):7220-8
pubmed: 11742998
Proc Natl Acad Sci U S A. 2014 Sep 16;111(37):E3900-9
pubmed: 25197083
J Virol. 2015 Apr;89(7):3598-609
pubmed: 25589656
Annu Rev Biochem. 2007;76:23-50
pubmed: 17506634
J Virol. 1999 Jul;73(7):5274-81
pubmed: 10364273
J Struct Biol. 2013 Nov;184(2):226-36
pubmed: 23954653
Nucleic Acids Res. 2003 Apr 1;31(7):1821-9
pubmed: 12654997
J Struct Biol X. 2019 Jan-Mar;1:
pubmed: 32285040
PLoS Pathog. 2011 Sep;7(9):e1002215
pubmed: 21931546
J Virol. 2000 Oct;74(20):9586-93
pubmed: 11000230
Cell. 2020 Jul 23;182(2):417-428.e13
pubmed: 32526208
Proc Natl Acad Sci U S A. 2012 Jun 12;109(24):9372-7
pubmed: 22635272
Virology. 2002 Jul 5;298(2):258-70
pubmed: 12127789