4CMQ

Crystal structure of Mn-bound S.pyogenes Cas9

4CMQ の概要

• エントリーDOI: 10.2210/pdb4cmq/pdb
• 関連するPDBエントリー: 4CMP
• 分子名称: CRISPR-ASSOCIATED ENDONUCLEASE CAS9/CSN1, MANGANESE (II) ION, SULFATE ION (3 entities in total)
• 機能のキーワード: hydrolase, dnase, endonuclease, rna-guided, immunity, crrna, genome editing
• 由来する生物種: STREPTOCOCCUS PYOGENES
• タンパク質・核酸の鎖数: 2
• 化学式量合計: 319262.79

構造登録者

Jinek, M.,Jiang, F.,Taylor, D.W.,Sternberg, S.H.,Kaya, E.,Ma, E.,Anders, C.,Hauer, M.,Zhou, K.,Lin, S.,Kaplan, M.,Iavarone, A.T.,Charpentier, E.,Nogales, E.,Doudna, J.A. (登録日: 2014-01-17, 公開日: 2014-02-12, 最終更新日: 2023-12-20)

主引用文献

Jinek, M.,Jiang, F.,Taylor, D.W.,Sternberg, S.H.,Kaya, E.,Ma, E.,Anders, C.,Hauer, M.,Zhou, K.,Lin, S.,Kaplan, M.,Iavarone, A.T.,Charpentier, E.,Nogales, E.,Doudna, J.A.
Structures of Cas9 Endonucleases Reveal RNA- Mediated Conformational Activation
Science, 343:47997-, 2014

PubMed Abstract: Type II CRISPR (clustered regularly interspaced short palindromic repeats)-Cas (CRISPR-associated) systems use an RNA-guided DNA endonuclease, Cas9, to generate double-strand breaks in invasive DNA during an adaptive bacterial immune response. Cas9 has been harnessed as a powerful tool for genome editing and gene regulation in many eukaryotic organisms. We report 2.6 and 2.2 angstrom resolution crystal structures of two major Cas9 enzyme subtypes, revealing the structural core shared by all Cas9 family members. The architectures of Cas9 enzymes define nucleic acid binding clefts, and single-particle electron microscopy reconstructions show that the two structural lobes harboring these clefts undergo guide RNA-induced reorientation to form a central channel where DNA substrates are bound. The observation that extensive structural rearrangements occur before target DNA duplex binding implicates guide RNA loading as a key step in Cas9 activation.

PubMed: 24505130
DOI: 10.1126/SCIENCE.1247997

実験手法

X-RAY DIFFRACTION (3.09 Å)