9UTM

Crystal structure of an anti-CRISPR protein AcrIIA11

9UTM の概要

• エントリーDOI: 10.2210/pdb9utm/pdb
• 分子名称: AcrIIA11 (1 entity in total)
• 機能のキーワード: cas9, anti-crispr, viral protein
• 由来する生物種: Clostridium sp. OF09-36
• タンパク質・核酸の鎖数: 2
• 化学式量合計: 44818.23

構造登録者

Wang, H.,Feng, Y. (登録日: 2025-05-04, 公開日: 2025-10-01)

主引用文献

Cao, X.,Gao, Z.,Yin, P.,Wang, H.,Yang, L.
Crystal structure and inhibition mechanism of AcrIIA11.
Biochem.Biophys.Res.Commun., 772:152073-152073, 2025

PubMed Abstract: Anti-CRISPR (Acr) proteins are naturally evolved inhibitors that precisely target and suppress CRISPR-Cas systems, representing a sophisticated molecular arms race between bacteriophages and their bacterial hosts. While Class 1 systems dominate among sequenced prokaryotic genomes, Class 2 systems remain primary sources of editing tools. Here, we report the structural and mechanistic characterization of AcrIIA11, an anti-CRISPR protein that simultaneously inhibits Streptococcus pyogenes (SpyCas9) and Staphylococcus aureus Cas9 (SauCas9). The 3.2 Å crystal structure reveals a compact α/β fold with distinct electropositive clefts implicated in DNA binding. While DALI analysis identified structural homology to transcriptional regulators and the RecA inhibitor PsiB (RMSD 3.3 Å), functional studies established that AcrIIA11 forms stable ternary complexes with both Cas9 orthologs and sgRNA. Biochemical assays demonstrated stronger inhibition of SauCas9 compared to SpyCas9, with EMSA revealing a critical dichotomy: AcrIIA11 maintains SauCas9-sgRNA binding to specific target DNA while completely blocking cleavage activity. Computational docking localizes AcrIIA11 at the HNH-RuvC interface without obstructing DNA-binding channels in SauCas9, suggesting allosteric inhibition through HNH domain displacement. This work establishes AcrIIA11 as a dual-purpose Cas9 inhibitor that preserves target recognition while inactivating nuclease function-a mechanism with potential applications in precision CRISPR control.

PubMed: 40414010
DOI: 10.1016/j.bbrc.2025.152073

実験手法

X-RAY DIFFRACTION (3.2 Å)