4ZT9

Nuclease-inactive Streptococcus pyogenes Cas9 (D10A/H840A, dCas9) in complex with single-guide RNA at 3.1 Angstrom resolution

4ZT9 の概要

• エントリーDOI: 10.2210/pdb4zt9/pdb
• 関連するPDBエントリー: 4ZT0
• 分子名称: CRISPR-associated endonuclease Cas9, single-guide RNA (2 entities in total)
• 機能のキーワード: crispr-cas9, bacteria adaptive immunity, genome editing and regulation, hydrolase-rna complex, hydrolase/rna
• 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: 372326.43

構造登録者

Jiang, F.,Doudna, J.A. (登録日: 2015-05-14, 公開日: 2015-07-08, 最終更新日: 2023-09-27)

主引用文献

Jiang, F.,Zhou, K.,Ma, L.,Gressel, S.,Doudna, J.A.
A Cas9-guide RNA complex preorganized for target DNA recognition.
Science, 348:1477-1481, 2015

PubMed Abstract: Bacterial adaptive immunity uses CRISPR (clustered regularly interspaced short palindromic repeats)-associated (Cas) proteins together with CRISPR transcripts for foreign DNA degradation. In type II CRISPR-Cas systems, activation of Cas9 endonuclease for DNA recognition upon guide RNA binding occurs by an unknown mechanism. Crystal structures of Cas9 bound to single-guide RNA reveal a conformation distinct from both the apo and DNA-bound states, in which the 10-nucleotide RNA "seed" sequence required for initial DNA interrogation is preordered in an A-form conformation. This segment of the guide RNA is essential for Cas9 to form a DNA recognition-competent structure that is poised to engage double-stranded DNA target sequences. We construe this as convergent evolution of a "seed" mechanism reminiscent of that used by Argonaute proteins during RNA interference in eukaryotes.

PubMed: 26113724
DOI: 10.1126/science.aab1452

実験手法

X-RAY DIFFRACTION (3.1 Å)