Structural basis of Staphylococcus aureus Cas9 inhibition by AcrIIA14.

Nucleic Acids Research
Hongnan LiuZhiwei Huang

Abstract

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.

References

Feb 18, 2014·Cell·Hiroshi NishimasuOsamu Nureki
Mar 22, 2014·Human Molecular Genetics·Feng ZhangXiong Guo
Apr 2, 2015·Nature·F Ann RanFeng Zhang
May 20, 2015·Methods in Molecular Biology·Kira S Makarova, Eugene V Koonin
Sep 1, 2015·Cell·Hiroshi NishimasuOsamu Nureki
Oct 17, 2015·Translational Research : the Journal of Laboratory and Clinical Medicine·Nataša Savić, Gerald Schwank
Nov 3, 2015·Nature·Samuel H SternbergJennifer A Doudna
Oct 5, 2016·Archivum Immunologiae Et Therapiae Experimentalis·Magdalena HryhorowiczRyszard Słomski
Apr 5, 2017·Annual Review of Biophysics·Fuguo Jiang, Jennifer A Doudna
Jun 13, 2017·Molecular Cell·Hui Yang, Dinshaw J Patel
Aug 16, 2017·Science Advances·Yavuz S DagdasAhmet Yildiz
Aug 29, 2017·Cell·Lucas B HarringtonJennifer A Doudna
Oct 7, 2017·Molecular Cell·Karen L Maxwell
Oct 25, 2017·Nature Reviews. Microbiology·April PawlukKaren L Maxwell
Nov 22, 2017·Trends in Biotechnology·Adrienne C Greene
Jan 24, 2018·Journal of Bacteriology·Yoshizumi IshinoPatrick Forterre
Apr 3, 2019·Nature Structural & Molecular Biology·Liyong DongZhiwei Huang
Apr 26, 2019·The CRISPR Journal·Sungwon Hwang, Karen L Maxwell
Jun 28, 2019·Nature Communications·Annoj ThavalingamKaren L Maxwell
Jul 10, 2019·Nature Structural & Molecular Biology·Xing ZhuSriram Subramaniam
Jul 12, 2019·Life Sciences·Darshana GuptaDipanjan Ghosh
Mar 12, 2020·Proceedings of the National Academy of Sciences of the United States of America·Kyle E WattersJennifer A Doudna
Mar 24, 2020·Nature Methods·Nicole D MarinoJoseph Bondy-Denomy
Apr 25, 2020·Nature Microbiology·Caroline MahendraJoseph Bondy-Denomy

❮ Previous
Next ❯

Software Mentioned

MOLPROBITY
Octet
PHASER
PHENIX
DALI
PROCHECK

Related Concepts

Related Feeds

Bacteriophage: Phage Therapy

Phage therapy uses bacterial viruses (bacteriophages) to treat bacterial infections and is widely being recognized as an alternative to antibiotics. Here is the latest research.

CRISPR Ribonucleases Deactivation

CRISPR-Cas system enables the editing of genes to create or correct mutations. This feed focuses on mechanisms that underlie deactivation of CRISPR ribonucleases. Here is the latest research.

CRISPR Genome Editing & Therapy

CRISPR-Cas system enables the editing of genes to create or correct mutations. This feed focuses on the application of this system for gene editing and therapy in human diseases.

CRISPR for Genome Editing

Genome editing technologies enable the editing of genes to create or correct mutations. Clustered regularly interspaced short palindromic repeats (CRISPR) are DNA sequences in the genome that are recognized and cleaved by CRISPR-associated proteins (Cas). Here is the latest research on the use of CRISPR-Cas system in gene editing.

CRISPR (general)

Clustered regularly interspaced short palindromic repeats (CRISPR) are DNA sequences in the genome that are recognized and cleaved by CRISPR-associated proteins (Cas). CRISPR-Cas system enables the editing of genes to create or correct mutations. Discover the latest research on CRISPR here.

Related Papers

BioRxiv : the Preprint Server for Biology
Kyle E WattersJennifer A Doudna
Proceedings of the National Academy of Sciences of the United States of America
Kyle E WattersJennifer A Doudna
© 2021 Meta ULC. All rights reserved