5F9R

Crystal structure of catalytically-active Streptococcus pyogenes CRISPR-Cas9 in complex with single-guided RNA and double-stranded DNA primed for target DNA cleavage

5F9R の概要

• エントリーDOI: 10.2210/pdb5f9r/pdb
• 分子名称: CRISPR-associated endonuclease Cas9/Csn1, RNA (116-MER), DNA (30-MER), ... (5 entities in total)
• 機能のキーワード: crispr, cas9, r-loop, genome engineering, hydrolase-dna-rna complex, hydrolase/dna/rna
• 由来する生物種: Streptococcus pyogenes serotype M1; 詳細
• タンパク質・核酸の鎖数: 4
• 化学式量合計: 215728.56

構造登録者

Jiang, F.,Doudna, J.A. (登録日: 2015-12-10, 公開日: 2016-01-27, 最終更新日: 2024-03-06)

主引用文献

Jiang, F.,Taylor, D.W.,Chen, J.S.,Kornfeld, J.E.,Zhou, K.,Thompson, A.J.,Nogales, E.,Doudna, J.A.
Structures of a CRISPR-Cas9 R-loop complex primed for DNA cleavage.
Science, 351:867-871, 2016

PubMed Abstract: Bacterial adaptive immunity and genome engineering involving the CRISPR (clustered regularly interspaced short palindromic repeats)-associated (Cas) protein Cas9 begin with RNA-guided DNA unwinding to form an RNA-DNA hybrid and a displaced DNA strand inside the protein. The role of this R-loop structure in positioning each DNA strand for cleavage by the two Cas9 nuclease domains is unknown. We determine molecular structures of the catalytically active Streptococcus pyogenes Cas9 R-loop that show the displaced DNA strand located near the RuvC nuclease domain active site. These protein-DNA interactions, in turn, position the HNH nuclease domain adjacent to the target DNA strand cleavage site in a conformation essential for concerted DNA cutting. Cas9 bends the DNA helix by 30°, providing the structural distortion needed for R-loop formation.

PubMed: 26841432
DOI: 10.1126/science.aad8282

実験手法

X-RAY DIFFRACTION (3.4 Å)