6LU0

Crystal structure of Cas12i2 ternary complex with 12 nt spacer

6LU0 の概要

• エントリーDOI: 10.2210/pdb6lu0/pdb
• 分子名称: Cas12i2, crRNA, DNA (5'-D(*GP*CP*CP*GP*CP*TP*TP*TP*CP*TP*T)-3'), ... (5 entities in total)
• 機能のキーワード: crispr-cas, cas12i2, cas12i2 binary complex, hydrolase, hydrolase-rna-dna complex, hydrolase/rna/dna
• 由来する生物種: unidentified; 詳細
• タンパク質・核酸の鎖数: 4
• 化学式量合計: 149882.89

構造登録者

Huang, X.,Sun, W.,Cheng, Z.,Chen, M.,Li, X.,Wang, J.,Sheng, G.,Gong, W.,Wang, Y. (登録日: 2020-01-24, 公開日: 2020-10-28, 最終更新日: 2023-11-29)

主引用文献

Huang, X.,Sun, W.,Cheng, Z.,Chen, M.,Li, X.,Wang, J.,Sheng, G.,Gong, W.,Wang, Y.
Structural basis for two metal-ion catalysis of DNA cleavage by Cas12i2.
Nat Commun, 11:5241-5241, 2020

PubMed Abstract: To understand how the RuvC catalytic domain of Class 2 Cas proteins cleaves DNA, it will be necessary to elucidate the structures of RuvC-containing Cas complexes in their catalytically competent states. Cas12i2 is a Class 2 type V-I CRISPR-Cas endonuclease that cleaves target dsDNA by an unknown mechanism. Here, we report structures of Cas12i2-crRNA-DNA complexes and a Cas12i2-crRNA complex. We reveal the mechanism of DNA recognition and cleavage by Cas12i2, and activation of the RuvC catalytic pocket induced by a conformational change of the Helical-II domain. The seed region (nucleotides 1-8) is dispensable for RuvC activation, but the duplex of the central spacer (nucleotides 9-15) is required. We captured the catalytic state of Cas12i2, with both metal ions and the ssDNA substrate bound in the RuvC catalytic pocket. Together, our studies provide significant insights into the DNA cleavage mechanism by RuvC-containing Cas proteins.

PubMed: 33067443
DOI: 10.1038/s41467-020-19072-6

実験手法

X-RAY DIFFRACTION (3.22 Å)