6IV8

the selenomethionine(SeMet)-derived Cas13d binary complex

6IV8 の概要

• エントリーDOI: 10.2210/pdb6iv8/pdb
• 分子名称: The selenomethionine (SeMet)-labeled Cas13d, RNA (51-MER), RNA (53-MER), ... (5 entities in total)
• 機能のキーワード: crispr, cas13d, crrna, binary complex, rna binding protein-rna complex, rna binding protein/rna
• 由来する生物種: uncultured Ruminococcus sp; 詳細
• タンパク質・核酸の鎖数: 4
• 化学式量合計: 250691.09

構造登録者

Zhang, B.,Ye, Y.M.,Ye, W.W.,OuYang, S.Y. (登録日: 2018-12-02, 公開日: 2019-06-19, 最終更新日: 2024-10-23)

主引用文献

Zhang, B.,Ye, Y.,Ye, W.,Perculija, V.,Jiang, H.,Chen, Y.,Li, Y.,Chen, J.,Lin, J.,Wang, S.,Chen, Q.,Han, Y.S.,Ouyang, S.
Two HEPN domains dictate CRISPR RNA maturation and target cleavage in Cas13d.
Nat Commun, 10:2544-2544, 2019

PubMed Abstract: Cas13d, the type VI-D CRISPR-Cas effector, is an RNA-guided ribonuclease that has been repurposed to edit RNA in a programmable manner. Here we report the detailed structural and functional analysis of the uncultured Ruminococcus sp. Cas13d (UrCas13d)-crRNA complex. Two hydrated Mg ions aid in stabilizing the conformation of the crRNA repeat region. Sequestration of divalent metal ions does not alter pre-crRNA processing, but abolishes target cleavage by UrCas13d. Notably, the pre-crRNA processing is executed by the HEPN-2 domain. Furthermore, both the structure and sequence of the nucleotides U(-8)-C(-1) within the repeat region are indispensable for target cleavage, and are specifically recognized by UrCas13d. Moreover, correct base pairings within two separate spacer regions (an internal and a 3'-end region) are essential for target cleavage. These findings provide a framework for the development of Cas13d into a tool for a wide range of applications.

PubMed: 31186424
DOI: 10.1038/s41467-019-10507-3

実験手法

X-RAY DIFFRACTION (2.15 Å)