5W1H

Crystal structure of LbaCas13a (C2c2) bound to mature crRNA (24-nt spacer)

5W1H の概要

• エントリーDOI: 10.2210/pdb5w1h/pdb
• 関連するPDBエントリー: 5W1I 5WLH
• 分子名称: LbaCas13a (C2c2), mature crRNA, IODIDE ION, ... (5 entities in total)
• 機能のキーワード: nuclease rna binding protein, rna binding protein, rna binding protein-rna complex, rna binding protein/rna
• 由来する生物種: Lachnospiraceae bacterium; 詳細
• タンパク質・核酸の鎖数: 2
• 化学式量合計: 189276.96

構造登録者

Knott, G.J.,Doudna, J.A. (登録日: 2017-06-03, 公開日: 2017-09-13, 最終更新日: 2024-03-13)

主引用文献

Knott, G.J.,East-Seletsky, A.,Cofsky, J.C.,Holton, J.M.,Charles, E.,O'Connell, M.R.,Doudna, J.A.
Guide-bound structures of an RNA-targeting A-cleaving CRISPR-Cas13a enzyme.
Nat. Struct. Mol. Biol., 24:825-833, 2017

PubMed Abstract: CRISPR adaptive immune systems protect bacteria from infections by deploying CRISPR RNA (crRNA)-guided enzymes to recognize and cut foreign nucleic acids. Type VI-A CRISPR-Cas systems include the Cas13a enzyme, an RNA-activated RNase capable of crRNA processing and single-stranded RNA degradation upon target-transcript binding. Here we present the 2.0-Å resolution crystal structure of a crRNA-bound Lachnospiraceae bacterium Cas13a (LbaCas13a), representing a recently discovered Cas13a enzyme subtype. This structure and accompanying biochemical experiments define the Cas13a catalytic residues that are directly responsible for crRNA maturation. In addition, the orientation of the foreign-derived target-RNA-specifying sequence in the protein interior explains the conformational gating of Cas13a nuclease activation. These results describe how Cas13a enzymes generate functional crRNAs and how catalytic activity is blocked before target-RNA recognition, with implications for both bacterial immunity and diagnostic applications.

PubMed: 28892041
DOI: 10.1038/nsmb.3466

実験手法

X-RAY DIFFRACTION (1.99 Å)