5U33

Crystal structure of AacC2c1-sgRNA-extended non-target DNA ternary complex

5U33 の概要

• エントリーDOI: 10.2210/pdb5u33/pdb
• 関連するPDBエントリー: 5U30 5U31 5U34
• 分子名称: CRISPR-associated endonuclease C2c1, sgRNA, Target DNA strand, ... (5 entities in total)
• 機能のキーワード: type v crispr-cas endonculease: c2c1: structure: binary complex with sgrna: ternary complex with added dna: ruvc catalytic pocket: sequence-specific pam recognition: genome editing tool, hydrolase-dna complex, hydrolase/dna
• 由来する生物種: Alicyclobacillus acidoterrestris (strain ATCC 49025 / DSM 3922 / CIP 106132 / NCIMB 13137 / GD3B); 詳細
• タンパク質・核酸の鎖数: 4
• 化学式量合計: 184106.79

構造登録者

Yang, H.,Gao, P.,Rajashankar, K.R.,Patel, D.J. (登録日: 2016-12-01, 公開日: 2017-01-25, 最終更新日: 2023-10-04)

主引用文献

Yang, H.,Gao, P.,Rajashankar, K.R.,Patel, D.J.
PAM-Dependent Target DNA Recognition and Cleavage by C2c1 CRISPR-Cas Endonuclease.
Cell, 167:1814-1828.e12, 2016

PubMed Abstract: C2c1 is a newly identified guide RNA-mediated type V-B CRISPR-Cas endonuclease that site-specifically targets and cleaves both strands of target DNA. We have determined crystal structures of Alicyclobacillus acidoterrestris C2c1 (AacC2c1) bound to sgRNA as a binary complex and to target DNAs as ternary complexes, thereby capturing catalytically competent conformations of AacC2c1 with both target and non-target DNA strands independently positioned within a single RuvC catalytic pocket. Moreover, C2c1-mediated cleavage results in a staggered seven-nucleotide break of target DNA. crRNA adopts a pre-ordered five-nucleotide A-form seed sequence in the binary complex, with release of an inserted tryptophan, facilitating zippering up of 20-bp guide RNA:target DNA heteroduplex on ternary complex formation. Notably, the PAM-interacting cleft adopts a "locked" conformation on ternary complex formation. Structural comparison of C2c1 ternary complexes with their Cas9 and Cpf1 counterparts highlights the diverse mechanisms adopted by these distinct CRISPR-Cas systems, thereby broadening and enhancing their applicability as genome editing tools.

PubMed: 27984729
DOI: 10.1016/j.cell.2016.11.053

実験手法

X-RAY DIFFRACTION (3.75 Å)