8D2Q

Structure of Acidothermus cellulolyticus Cas9 ternary complex (Post-cleavage 1)

8D2Q の概要

• エントリーDOI: 10.2210/pdb8d2q/pdb
• 関連するPDBエントリー: 8D2P
• EMDBエントリー: 27145 27146
• 分子名称: CRISPR-associated endonuclease, Csn1 family, Single Guide RNA (86-MER), DNA target strand (5'-D(P*CP*CP*AP*GP*GP*AP*TP*CP*TP*TP*G)-3'), ... (7 entities in total)
• 機能のキーワード: cas9, acecas9, crispr, post-cleavage 1, rna binding protein, rna binding protein-dna-rna complex, rna binding protein/dna/rna
• 由来する生物種: Acidothermus cellulolyticus 11B; 詳細
• タンパク質・核酸の鎖数: 5
• 化学式量合計: 172996.13

構造登録者

Rai, J.,Das, A.,Li, H. (登録日: 2022-05-30, 公開日: 2023-12-20, 最終更新日: 2025-05-28)

主引用文献

Das, A.,Rai, J.,Roth, M.O.,Shu, Y.,Medina, M.L.,Barakat, M.R.,Li, H.
Coupled catalytic states and the role of metal coordination in Cas9.
Nat Catal, 6:969-977, 2023

PubMed Abstract: Controlling the activity of the CRISPR-Cas9 system is essential to its safe adoption for clinical and research applications. Although the conformational dynamics of Cas9 are known to control its enzymatic activity, details of how Cas9 influences the catalytic processes at both nuclease domains remain elusive. Here we report five cryo-electron microscopy structures of the active Cas9 complex along the reaction path at 2.2-2.9 Å resolution. We observed that a large movement in one nuclease domain, triggered by the cognate DNA, results in noticeable changes in the active site of the other domain that is required for metal coordination and catalysis. Furthermore, the conformations synchronize the reaction intermediates, enabling coupled cutting of the two DNA strands. Consistent with the roles of conformations in organizing the active sites, adjustments to the metal-coordination residues lead to altered metal specificity of Cas9 and commonly used Cas9 in cells.

PubMed: 38348449
DOI: 10.1038/s41929-023-01031-1

実験手法

ELECTRON MICROSCOPY (2.58 Å)