6MCC

CryoEM structure of AcrIIA2 homolog in complex with CRISPR-Cas9

6MCC の概要

• エントリーDOI: 10.2210/pdb6mcc/pdb
• EMDBエントリー: 9066 9067
• 分子名称: Single guide RNA (116-MER), CRISPR-associated endonuclease Cas9, Anti-CRISPR AcrIIA2 Homolog (3 entities in total)
• 機能のキーワード: crispr-cas, cas9, cas9 inhibitors, anti-crispr, acriia2, bacteriophage, gene editing, hydrolase-rna-viral protein complex, hydrolase/rna/viral protein
• 由来する生物種: Streptococcus pyogenes M1 GAS; 詳細
• タンパク質・核酸の鎖数: 3
• 化学式量合計: 211625.77

構造登録者

Jiang, F.,Liu, J.J.,Doudna, J.A. (登録日: 2018-08-31, 公開日: 2019-01-16, 最終更新日: 2024-03-13)

主引用文献

Jiang, F.,Liu, J.J.,Osuna, B.A.,Xu, M.,Berry, J.D.,Rauch, B.J.,Nogales, E.,Bondy-Denomy, J.,Doudna, J.A.
Temperature-Responsive Competitive Inhibition of CRISPR-Cas9.
Mol. Cell, 73:601-, 2019

PubMed Abstract: CRISPR-Cas immune systems utilize RNA-guided nucleases to protect bacteria from bacteriophage infection. Bacteriophages have in turn evolved inhibitory "anti-CRISPR" (Acr) proteins, including six inhibitors (AcrIIA1-AcrIIA6) that can block DNA cutting and genome editing by type II-A CRISPR-Cas9 enzymes. We show here that AcrIIA2 and its more potent homolog, AcrIIA2b, prevent Cas9 binding to DNA by occluding protein residues required for DNA binding. Cryo-EM-determined structures of AcrIIA2 or AcrIIA2b bound to S. pyogenes Cas9 reveal a mode of competitive inhibition of DNA binding that is distinct from other known Acrs. Differences in the temperature dependence of Cas9 inhibition by AcrIIA2 and AcrIIA2b arise from differences in both inhibitor structure and the local inhibitor-binding environment on Cas9. These findings expand the natural toolbox for regulating CRISPR-Cas9 genome editing temporally, spatially, and conditionally.

PubMed: 30595438
DOI: 10.1016/j.molcel.2018.11.016

実験手法

ELECTRON MICROSCOPY (3.9 Å)