7V93

Cryo-EM structure of the Cas12c2-sgRNA binary complex

これはPDB形式変換不可エントリーです。

7V93 の概要

• エントリーDOI: 10.2210/pdb7v93/pdb
• EMDBエントリー: 31807 31808
• 分子名称: cas12c2, sgRNA (2 entities in total)
• 機能のキーワード: cas12c, c2c3, sgrna, crispr, rna binding protein-rna complex, rna binding protein/rna
• 由来する生物種: uncultured archaeon; 詳細
• タンパク質・核酸の鎖数: 2
• 化学式量合計: 174496.35

構造登録者

Kurihara, N.,Hirano, H.,Tomita, A.,Kobayashi, K.,Kusakizako, T.,Nishizawa, T.,Yamashita, K.,Nishimasu, H.,Nureki, O. (登録日: 2021-08-24, 公開日: 2022-04-13, 最終更新日: 2024-06-12)

主引用文献

Kurihara, N.,Nakagawa, R.,Hirano, H.,Okazaki, S.,Tomita, A.,Kobayashi, K.,Kusakizako, T.,Nishizawa, T.,Yamashita, K.,Scott, D.A.,Nishimasu, H.,Nureki, O.
Structure of the type V-C CRISPR-Cas effector enzyme.
Mol.Cell, 82:1865-1877.e4, 2022

PubMed Abstract: RNA-guided CRISPR-Cas nucleases are widely used as versatile genome-engineering tools. Recent studies identified functionally divergent type V Cas12 family enzymes. Among them, Cas12c2 binds a CRISPR RNA (crRNA) and a trans-activating crRNA (tracrRNA) and recognizes double-stranded DNA targets with a short TN PAM. Here, we report the cryo-electron microscopy structures of the Cas12c2-guide RNA binary complex and the Cas12c2-guide RNA-target DNA ternary complex. The structures revealed that the crRNA and tracrRNA form an unexpected X-junction architecture, and that Cas12c2 recognizes a single T nucleotide in the PAM through specific hydrogen-bonding interactions with two arginine residues. Furthermore, our biochemical analyses indicated that Cas12c2 processes its precursor crRNA to a mature crRNA using the RuvC catalytic site through a unique mechanism. Collectively, our findings improve the mechanistic understanding of diverse type V CRISPR-Cas effectors.

PubMed: 35366394
DOI: 10.1016/j.molcel.2022.03.006

実験手法

ELECTRON MICROSCOPY (3 Å)