7XG0

CryoEM structure of type IV-A Csf-crRNA-dsDNA ternary complex

7XG0 の概要

• エントリーDOI: 10.2210/pdb7xg0/pdb
• EMDBエントリー: 33181
• 分子名称: Csf1, Csf3, Csf2, ... (8 entities in total)
• 機能のキーワード: nuclease, structural protein-rna-dna complex, structural protein/rna/dna
• 由来する生物種: Pseudomonas aeruginosa; 詳細
• タンパク質・核酸の鎖数: 11
• 化学式量合計: 322104.84

構造登録者

Zhang, J.T.,Cui, N.,Huang, H.D.,Jia, N. (登録日: 2022-04-02, 公開日: 2023-08-09, 最終更新日: 2024-07-03)

主引用文献

Cui, N.,Zhang, J.T.,Liu, Y.,Liu, Y.,Liu, X.Y.,Wang, C.,Huang, H.,Jia, N.
Type IV-A CRISPR-Csf complex: Assembly, dsDNA targeting, and CasDinG recruitment.
Mol.Cell, 83:2493-2508.e5, 2023

PubMed Abstract: Type IV CRISPR-Cas systems, which are primarily found on plasmids and exhibit a strong plasmid-targeting preference, are the only one of the six known CRISPR-Cas types for which the mechanistic details of their function remain unknown. Here, we provide high-resolution functional snapshots of type IV-A Csf complexes before and after target dsDNA binding, either in the absence or presence of CasDinG, revealing the mechanisms underlying Csf complex assembly, "DWN" PAM-dependent dsDNA targeting, R-loop formation, and CasDinG recruitment. Furthermore, we establish that CasDinG, a signature DinG family helicase, harbors ssDNA-stimulated ATPase activity and ATP-dependent 5'-3' DNA helicase activity. In addition, we show that CasDinG unwinds the non-target strand (NTS) and target strand (TS) of target dsDNA from the Csf complex. These molecular details advance our mechanistic understanding of type IV-A CRISPR-Csf function and should enable Csf complexes to be harnessed as genome-engineering tools for biotechnological applications.

PubMed: 37343553
DOI: 10.1016/j.molcel.2023.05.036

実験手法

ELECTRON MICROSCOPY (2.6 Å)