5B43

Crystal structure of Acidaminococcus sp. Cpf1 in complex with crRNA and target DNA

5B43 の概要

• エントリーDOI: 10.2210/pdb5b43/pdb
• 分子名称: CRISPR-associated endonuclease Cpf1, RNA (43-MER), DNA (34-MER), ... (7 entities in total)
• 機能のキーワード: nuclease, hydrolase-rna-dna complex, hydrolase/rna/dna
• 由来する生物種: Acidaminococcus sp. BV3L6; 詳細
• タンパク質・核酸の鎖数: 4
• 化学式量合計: 179105.26

構造登録者

Yamano, T.,Nishimasu, H.,Hirano, H.,Nakane, T.,Ishitani, R.,Nureki, O. (登録日: 2016-03-30, 公開日: 2016-05-04, 最終更新日: 2024-03-20)

主引用文献

Yamano, T.,Nishimasu, H.,Zetsche, B.,Hirano, H.,Slaymaker, I.M.,Li, Y.,Fedorova, I.,Nakane, T.,Makarova, K.S.,Koonin, E.V.,Ishitani, R.,Zhang, F.,Nureki, O.
Crystal Structure of Cpf1 in Complex with Guide RNA and Target DNA
Cell, 165:949-962, 2016

PubMed Abstract: Cpf1 is an RNA-guided endonuclease of a type V CRISPR-Cas system that has been recently harnessed for genome editing. Here, we report the crystal structure of Acidaminococcus sp. Cpf1 (AsCpf1) in complex with the guide RNA and its target DNA at 2.8 Å resolution. AsCpf1 adopts a bilobed architecture, with the RNA-DNA heteroduplex bound inside the central channel. The structural comparison of AsCpf1 with Cas9, a type II CRISPR-Cas nuclease, reveals both striking similarity and major differences, thereby explaining their distinct functionalities. AsCpf1 contains the RuvC domain and a putative novel nuclease domain, which are responsible for cleaving the non-target and target strands, respectively, and for jointly generating staggered DNA double-strand breaks. AsCpf1 recognizes the 5'-TTTN-3' protospacer adjacent motif by base and shape readout mechanisms. Our findings provide mechanistic insights into RNA-guided DNA cleavage by Cpf1 and establish a framework for rational engineering of the CRISPR-Cpf1 toolbox.

PubMed: 27114038
DOI: 10.1016/j.cell.2016.04.003

実験手法

X-RAY DIFFRACTION (2.8 Å)