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Structural basis of Staphylococcus aureus Cas9 inhibition by AcrIIA14
Bacteriophages have evolved a range of anti-CRISPR proteins (Acrs) to escape the adaptive immune system of prokaryotes, therefore Acrs can be used as switches to regulate gene editing. Herein, we report the crystal structure of a quaternary complex of AcrIIA14 bound SauCas9–sgRNA–dsDNA at 2.22 Å res...
Autores principales: | , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Oxford University Press
2021
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8216286/ https://www.ncbi.nlm.nih.gov/pubmed/34107040 http://dx.doi.org/10.1093/nar/gkab487 |
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author | Liu, Hongnan Zhu, Yuwei Lu, Zebin Huang, Zhiwei |
author_facet | Liu, Hongnan Zhu, Yuwei Lu, Zebin Huang, Zhiwei |
author_sort | Liu, Hongnan |
collection | PubMed |
description | Bacteriophages have evolved a range of anti-CRISPR proteins (Acrs) to escape the adaptive immune system of prokaryotes, therefore Acrs can be used as switches to regulate gene editing. Herein, we report the crystal structure of a quaternary complex of AcrIIA14 bound SauCas9–sgRNA–dsDNA at 2.22 Å resolution, revealing the molecular basis for AcrIIA14 recognition and inhibition. Our structural and biochemical data analysis suggest that AcrIIA14 binds to a non-conserved region of SauCas9 HNH domain that is distinctly different from AcrIIC1 and AcrIIC3, with no significant effect on sgRNA or dsDNA binding. Further, our structural data shows that the allostery of the HNH domain close to the substrate DNA is sterically prevented by AcrIIA14 binding. In addition, the binding of AcrIIA14 triggers the conformational allostery of the HNH domain and the L1 linker within the SauCas9, driving them to make new interactions with the target-guide heteroduplex, enhancing the inhibitory ability of AcrIIA14. Our research both expands the current understanding of anti-CRISPRs and provides additional culues for the rational use of the CRISPR-Cas system in genome editing and gene regulation. |
format | Online Article Text |
id | pubmed-8216286 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2021 |
publisher | Oxford University Press |
record_format | MEDLINE/PubMed |
spelling | pubmed-82162862021-06-22 Structural basis of Staphylococcus aureus Cas9 inhibition by AcrIIA14 Liu, Hongnan Zhu, Yuwei Lu, Zebin Huang, Zhiwei Nucleic Acids Res Structural Biology Bacteriophages have evolved a range of anti-CRISPR proteins (Acrs) to escape the adaptive immune system of prokaryotes, therefore Acrs can be used as switches to regulate gene editing. Herein, we report the crystal structure of a quaternary complex of AcrIIA14 bound SauCas9–sgRNA–dsDNA at 2.22 Å resolution, revealing the molecular basis for AcrIIA14 recognition and inhibition. Our structural and biochemical data analysis suggest that AcrIIA14 binds to a non-conserved region of SauCas9 HNH domain that is distinctly different from AcrIIC1 and AcrIIC3, with no significant effect on sgRNA or dsDNA binding. Further, our structural data shows that the allostery of the HNH domain close to the substrate DNA is sterically prevented by AcrIIA14 binding. In addition, the binding of AcrIIA14 triggers the conformational allostery of the HNH domain and the L1 linker within the SauCas9, driving them to make new interactions with the target-guide heteroduplex, enhancing the inhibitory ability of AcrIIA14. Our research both expands the current understanding of anti-CRISPRs and provides additional culues for the rational use of the CRISPR-Cas system in genome editing and gene regulation. Oxford University Press 2021-06-09 /pmc/articles/PMC8216286/ /pubmed/34107040 http://dx.doi.org/10.1093/nar/gkab487 Text en © The Author(s) 2021. Published by Oxford University Press on behalf of Nucleic Acids Research. https://creativecommons.org/licenses/by/4.0/This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0/ (https://creativecommons.org/licenses/by/4.0/) ), which permits unrestricted reuse, distribution, and reproduction in any medium, provided the original work is properly cited. |
spellingShingle | Structural Biology Liu, Hongnan Zhu, Yuwei Lu, Zebin Huang, Zhiwei Structural basis of Staphylococcus aureus Cas9 inhibition by AcrIIA14 |
title | Structural basis of Staphylococcus aureus Cas9 inhibition by AcrIIA14 |
title_full | Structural basis of Staphylococcus aureus Cas9 inhibition by AcrIIA14 |
title_fullStr | Structural basis of Staphylococcus aureus Cas9 inhibition by AcrIIA14 |
title_full_unstemmed | Structural basis of Staphylococcus aureus Cas9 inhibition by AcrIIA14 |
title_short | Structural basis of Staphylococcus aureus Cas9 inhibition by AcrIIA14 |
title_sort | structural basis of staphylococcus aureus cas9 inhibition by acriia14 |
topic | Structural Biology |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8216286/ https://www.ncbi.nlm.nih.gov/pubmed/34107040 http://dx.doi.org/10.1093/nar/gkab487 |
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