7TRA

Cascade complex from type I-A CRISPR-Cas system

これはPDB形式変換不可エントリーです。

7TRA の概要

• エントリーDOI: 10.2210/pdb7tra/pdb
• EMDBエントリー: 26084
• 分子名称: CRISPR-associated helicase Cas3, NICKEL (II) ION, Cas8a, ... (10 entities in total)
• 機能のキーワード: crispr, cascade, cas3, type i-a, genome editing, dna targeting, rna binding protein
• 由来する生物種: Pyrococcus furiosus DSM 3638; 詳細
• タンパク質・核酸の鎖数: 19
• 化学式量合計: 509673.02

構造登録者

Hu, C.,Ni, D.,Nam, K.H.,Majumdar, S.,McLean, J.,Stahlberg, H.,Terns, M.,Ke, A. (登録日: 2022-01-28, 公開日: 2022-08-10, 最終更新日: 2025-05-14)

主引用文献

Hu, C.,Ni, D.,Nam, K.H.,Majumdar, S.,McLean, J.,Stahlberg, H.,Terns, M.P.,Ke, A.
Allosteric control of type I-A CRISPR-Cas3 complexes and establishment as effective nucleic acid detection and human genome editing tools.
Mol.Cell, 82:2754-2768.e5, 2022

PubMed Abstract: Type I CRISPR-Cas systems typically rely on a two-step process to degrade DNA. First, an RNA-guided complex named Cascade identifies the complementary DNA target. The helicase-nuclease fusion enzyme Cas3 is then recruited in trans for processive DNA degradation. Contrary to this model, here, we show that type I-A Cascade and Cas3 function as an integral effector complex. We provide four cryoelectron microscopy (cryo-EM) snapshots of the Pyrococcus furiosus (Pfu) type I-A effector complex in different stages of DNA recognition and degradation. The HD nuclease of Cas3 is autoinhibited inside the effector complex. It is only allosterically activated upon full R-loop formation, when the entire targeted region has been validated by the RNA guide. The mechanistic insights inspired us to convert Pfu Cascade-Cas3 into a high-sensitivity, low-background, and temperature-activated nucleic acid detection tool. Moreover, Pfu CRISPR-Cas3 shows robust bi-directional deletion-editing activity in human cells, which could find usage in allele-specific inactivation of disease-causing mutations.

PubMed: 35835111
DOI: 10.1016/j.molcel.2022.06.007

実験手法

ELECTRON MICROSCOPY (3.3 Å)