6O0Z

Conformational states of Cas9-sgRNA-DNA ternary complex in the presence of magnesium

6O0Z の概要

• エントリーDOI: 10.2210/pdb6o0z/pdb
• EMDBエントリー: 0585
• 分子名称: CRISPR-associated endonuclease Cas9/Csn1, single guide RNA, target strand DNA, ... (4 entities in total)
• 機能のキーワード: crispr, cas9, nuclease, genome editing, hydrolase, hydrolase-rna-dna complex, hydrolase/rna/dna
• 由来する生物種: Streptococcus pyogenes; 詳細
• タンパク質・核酸の鎖数: 4
• 化学式量合計: 216673.61

構造登録者

Zhu, X.,Clarke, R.,Puppala, A.K.,Chittori, S.,Merk, A.,Merrill, B.J.,Simonovic, M.,Subramaniam, S. (登録日: 2019-02-17, 公開日: 2019-07-10, 最終更新日: 2025-05-21)

主引用文献

Zhu, X.,Clarke, R.,Puppala, A.K.,Chittori, S.,Merk, A.,Merrill, B.J.,Simonovic, M.,Subramaniam, S.
Cryo-EM structures reveal coordinated domain motions that govern DNA cleavage by Cas9.
Nat.Struct.Mol.Biol., 26:679-685, 2019

PubMed Abstract: The RNA-guided Cas9 endonuclease from Streptococcus pyogenes is a single-turnover enzyme that displays a stable product state after double-stranded-DNA cleavage. Here, we present cryo-EM structures of precatalytic, postcatalytic and product states of the active Cas9-sgRNA-DNA complex in the presence of Mg. In the precatalytic state, Cas9 adopts the 'checkpoint' conformation with the HNH nuclease domain positioned far away from the DNA. Transition to the postcatalytic state involves a dramatic ~34-Å swing of the HNH domain and disorder of the REC2 recognition domain. The postcatalytic state captures the cleaved substrate bound to the catalytically competent HNH active site. In the product state, the HNH domain is disordered, REC2 returns to the precatalytic conformation, and additional interactions of REC3 and RuvC with nucleic acids are formed. The coupled domain motions and interactions between the enzyme and the RNA-DNA hybrid provide new insights into the mechanism of genome editing by Cas9.

PubMed: 31285607
DOI: 10.1038/s41594-019-0258-2

実験手法

ELECTRON MICROSCOPY (3.3 Å)