5CD4

The Type IE CRISPR Cascade complex from E. coli, with two assemblies in the asymmetric unit arranged back-to-back

5CD4 の概要

• エントリーDOI: 10.2210/pdb5cd4/pdb
• 分子名称: CRISPR system Cascade subunit CasE, CRISPR system Cascade subunit CasC, CRISPR system Cascade subunit CasD, ... (7 entities in total)
• 機能のキーワード: crispr, cascade, rna surveillance, adaptive immunity, hydrolase-rna complex, hydrolase/rna
• 由来する生物種: Escherichia coli; 詳細
• タンパク質・核酸の鎖数: 24
• 化学式量合計: 804324.65

構造登録者

Jackson, R.N.,Golden, S.M.,Carter, J.,Wiedenheft, B. (登録日: 2015-07-03, 公開日: 2015-08-26, 最終更新日: 2024-12-25)

主引用文献

van Erp, P.B.,Jackson, R.N.,Carter, J.,Golden, S.M.,Bailey, S.,Wiedenheft, B.
Mechanism of CRISPR-RNA guided recognition of DNA targets in Escherichia coli.
Nucleic Acids Res., 43:8381-8391, 2015

PubMed Abstract: In bacteria and archaea, short fragments of foreign DNA are integrated into Clustered Regularly Interspaced Short Palindromic Repeat (CRISPR) loci, providing a molecular memory of previous encounters with foreign genetic elements. In Escherichia coli, short CRISPR-derived RNAs are incorporated into a multi-subunit surveillance complex called Cascade (CRISPR-associated complex for antiviral defense). Recent structures of Cascade capture snapshots of this seahorse-shaped RNA-guided surveillance complex before and after binding to a DNA target. Here we determine a 3.2 Å x-ray crystal structure of Cascade in a new crystal form that provides insight into the mechanism of double-stranded DNA binding. Molecular dynamic simulations performed using available structures reveal functional roles for residues in the tail, backbone and belly subunits of Cascade that are critical for binding double-stranded DNA. Structural comparisons are used to make functional predictions and these predictions are tested in vivo and in vitro. Collectively, the results in this study reveal underlying mechanisms involved in target-induced conformational changes and highlight residues important in DNA binding and protospacer adjacent motif recognition.

PubMed: 26243775
DOI: 10.1093/nar/gkv793

実験手法

X-RAY DIFFRACTION (3.2 Å)