8JA0

Cryo-EM structure of the NmeCas9-sgRNA-AcrIIC4 ternary complex

8JA0 の概要

• エントリーDOI: 10.2210/pdb8ja0/pdb
• EMDBエントリー: 36123
• 分子名称: CRISPR-associated endonuclease Cas9, Uncharacterized protein, RNA (117-MER) (3 entities in total)
• 機能のキーワード: a protein complex, viral protein, rna binding protein-rna complex, rna binding protein/rna
• 由来する生物種: Neisseria meningitidis; 詳細
• タンパク質・核酸の鎖数: 3
• 化学式量合計: 171966.26

構造登録者

Yin, H.,Li, Z.,Yu, G.M.,Li, X.Z. (登録日: 2023-05-05, 公開日: 2023-08-09, 最終更新日: 2025-07-02)

主引用文献

Li, X.,Liao, F.,Gao, J.,Song, G.,Zhang, C.,Ji, N.,Wang, X.,Wen, J.,He, J.,Wei, Y.,Zhang, H.,Li, Z.,Yu, G.,Yin, H.
Inhibitory mechanism of CRISPR-Cas9 by AcrIIC4.
Nucleic Acids Res., 51:9442-9451, 2023

PubMed Abstract: CRISPR-Cas systems act as the adaptive immune systems of bacteria and archaea, targeting and destroying invading foreign mobile genetic elements (MGEs) such as phages. MGEs have also evolved anti-CRISPR (Acr) proteins to inactivate the CRISPR-Cas systems. Recently, AcrIIC4, identified from Haemophilus parainfluenzae phage, has been reported to inhibit the endonuclease activity of Cas9 from Neisseria meningitidis (NmeCas9), but the inhibition mechanism is not clear. Here, we biochemically and structurally investigated the anti-CRISPR activity of AcrIIC4. AcrIIC4 folds into a helix bundle composed of three helices, which associates with the REC lobe of NmeCas9 and sgRNA. The REC2 domain of NmeCas9 is locked by AcrIIC4, perturbing the conformational dynamics required for the target DNA binding and cleavage. Furthermore, mutation of the key residues in the AcrIIC4-NmeCas9 and AcrIIC4-sgRNA interfaces largely abolishes the inhibitory effects of AcrIIC4. Our study offers new insights into the mechanism of AcrIIC4-mediated suppression of NmeCas9 and provides guidelines for the design of regulatory tools for Cas9-based gene editing applications.

PubMed: 37587688
DOI: 10.1093/nar/gkad669

実験手法

ELECTRON MICROSCOPY (3.52 Å)