Anti-CRISPR AcrIIC5 is a dsDNA mimic that inhibits type II-C Cas9 effectors by blocking PAM recognition.
Journal
Nucleic acids research
ISSN: 1362-4962
Titre abrégé: Nucleic Acids Res
Pays: England
ID NLM: 0411011
Informations de publication
Date de publication:
28 02 2023
28 02 2023
Historique:
accepted:
21
01
2023
revised:
16
01
2023
received:
28
11
2022
pubmed:
7
2
2023
medline:
4
3
2023
entrez:
6
2
2023
Statut:
ppublish
Résumé
Anti-CRISPR proteins are encoded by phages to inhibit the CRISPR-Cas systems of the hosts. AcrIIC5 inhibits several naturally high-fidelity type II-C Cas9 enzymes, including orthologs from Neisseria meningitidis (Nme1Cas9) and Simonsiella muelleri (SmuCas9). Here, we solve the structure of AcrIIC5 in complex with Nme1Cas9 and sgRNA. We show that AcrIIC5 adopts a novel fold to mimic the size and charge distribution of double-stranded DNA, and uses its negatively charged grooves to bind and occlude the protospacer adjacent motif (PAM) binding site in the target DNA cleft of Cas9. AcrIIC5 is positioned into the crevice between the WED and PI domains of Cas9, and one end of the anti-CRISPR interacts with the phosphate lock loop and a linker between the RuvC and BH domains. We employ biochemical and mutational analyses to build a model for AcrIIC5's mechanism of action, and identify residues on both the anti-CRISPR and Cas9 that are important for their interaction and inhibition. Together, the structure and mechanism of AcrIIC5 reveal convergent evolution among disparate anti-CRISPR proteins that use a DNA-mimic strategy to inhibit diverse CRISPR-Cas surveillance complexes, and provide new insights into a tool for potent inhibition of type II-C Cas9 orthologs.
Identifiants
pubmed: 36744495
pii: 7028258
doi: 10.1093/nar/gkad052
pmc: PMC9976890
doi:
Substances chimiques
CRISPR-Associated Protein 9
EC 3.1.-
DNA
9007-49-2
Viral Proteins
0
Types de publication
Journal Article
Research Support, Non-U.S. Gov't
Langues
eng
Sous-ensembles de citation
IM
Pagination
1984-1995Informations de copyright
© The Author(s) 2023. Published by Oxford University Press on behalf of Nucleic Acids Research.
Références
Mol Cell. 2019 Apr 18;74(2):296-309.e7
pubmed: 30850331
J Comput Chem. 2004 Oct;25(13):1605-12
pubmed: 15264254
Nat Methods. 2017 Mar;14(3):290-296
pubmed: 28165473
Nature. 2020 Jan;577(7791):572-575
pubmed: 31942067
Mol Cell. 2019 Feb 7;73(3):611-620.e3
pubmed: 30606466
Nature. 2021 Aug;596(7873):583-589
pubmed: 34265844
Mol Cell. 2017 Oct 5;68(1):8-14
pubmed: 28985512
Nat Rev Microbiol. 2020 Feb;18(2):67-83
pubmed: 31857715
Nucleic Acids Res. 2011 Nov;39(21):9275-82
pubmed: 21813460
Science. 2020 Jul 3;369(6499):54-59
pubmed: 32467331
Nature. 2011 Mar 31;471(7340):602-7
pubmed: 21455174
Mol Cell. 2014 Apr 24;54(2):234-44
pubmed: 24766887
Cell. 2016 Jan 14;164(1-2):29-44
pubmed: 26771484
Proc Natl Acad Sci U S A. 2013 Sep 24;110(39):15644-9
pubmed: 23940360
Cell. 2017 Jan 12;168(1-2):150-158.e10
pubmed: 28041849
Nature. 2017 Jun 15;546(7658):436-439
pubmed: 28448066
Nature. 2014 Sep 25;513(7519):569-73
pubmed: 25079318
Genome Biol. 2018 Dec 5;19(1):214
pubmed: 30518407
ACS Chem Biol. 2018 Feb 16;13(2):357-365
pubmed: 29202216
Science. 2008 Dec 19;322(5909):1843-5
pubmed: 19095942
Science. 2016 Feb 19;351(6275):867-71
pubmed: 26841432
Nature. 2013 Jan 17;493(7432):429-32
pubmed: 23242138
Nat Commun. 2019 Jun 26;10(1):2806
pubmed: 31243272
Annu Rev Biophys. 2017 May 22;46:505-529
pubmed: 28375731
Science. 2007 Mar 23;315(5819):1709-12
pubmed: 17379808
Cell. 2017 Oct 5;171(2):414-426.e12
pubmed: 28985564
Mol Cell. 2019 Dec 19;76(6):938-952.e5
pubmed: 31668930
Cell. 2017 Sep 7;170(6):1224-1233.e15
pubmed: 28844692
Mol Cell. 2017 Mar 16;65(6):1109-1121.e3
pubmed: 28306506
Nucleic Acids Res. 2019 Jul 26;47(13):e75
pubmed: 30982889
Protein Sci. 2021 Jan;30(1):70-82
pubmed: 32881101
Acta Crystallogr D Biol Crystallogr. 2010 Apr;66(Pt 4):486-501
pubmed: 20383002
Annu Rev Biochem. 2020 Jun 20;89:309-332
pubmed: 32186918
Mol Cell. 2019 Feb 7;73(3):601-610.e5
pubmed: 30595438
Mol Cell. 2017 Jul 6;67(1):117-127.e5
pubmed: 28602637
Mol Cell. 2019 Feb 21;73(4):714-726.e4
pubmed: 30581144
Cell. 2016 Dec 15;167(7):1829-1838.e9
pubmed: 27984730
Cell. 2017 Mar 23;169(1):47-57.e11
pubmed: 28340349
Sci Adv. 2017 Jul 12;3(7):e1701620
pubmed: 28706995
Acta Crystallogr D Biol Crystallogr. 2002 Nov;58(Pt 11):1948-54
pubmed: 12393927
Science. 2008 Aug 15;321(5891):960-4
pubmed: 18703739
Science. 2018 Oct 12;362(6411):240-242
pubmed: 30190308
Nature. 2010 Nov 4;468(7320):67-71
pubmed: 21048762
mBio. 2018 Dec 4;9(6):
pubmed: 30514786
Mol Cell. 2022 Jan 20;82(2):333-347
pubmed: 34968414
Annu Rev Microbiol. 2020 Sep 8;74:21-37
pubmed: 32503371
RNA. 2019 Nov;25(11):1421-1431
pubmed: 31439808
Science. 2018 Oct 12;362(6411):236-239
pubmed: 30190307
Mol Cell. 2019 Dec 19;76(6):922-937.e7
pubmed: 31604602
Cell. 2017 Jan 12;168(1-2):20-36
pubmed: 27866654