8GS2

Structure of the Cas7-11-Csx29-guide RNA-target RNA (non-matching PFS) complex

8GS2 の概要

• エントリーDOI: 10.2210/pdb8gs2/pdb
• EMDBエントリー: 34218
• 分子名称: crRNA, target RNA with non-matching PFS, CRISPR-associated RAMP family protein, ... (7 entities in total)
• 機能のキーワード: crispr, rnase, rna binding protein-rna complex, rna binding protein/rna
• 由来する生物種: Desulfonema ishimotonii; 詳細
• タンパク質・核酸の鎖数: 4
• 化学式量合計: 316390.27

構造登録者

Kato, K.,Okazaki, S.,Ishikawa, J.,Isayama, Y.,Nishizawa, T.,Nishimasu, H. (登録日: 2022-09-04, 公開日: 2022-11-09, 最終更新日: 2024-05-29)

主引用文献

Kato, K.,Okazaki, S.,Schmitt-Ulms, C.,Jiang, K.,Zhou, W.,Ishikawa, J.,Isayama, Y.,Adachi, S.,Nishizawa, T.,Makarova, K.S.,Koonin, E.V.,Abudayyeh, O.O.,Gootenberg, J.S.,Nishimasu, H.
RNA-triggered protein cleavage and cell growth arrest by the type III-E CRISPR nuclease-protease.
Science, 378:882-889, 2022

PubMed Abstract: The type III-E CRISPR-Cas7-11 effector binds a CRISPR RNA (crRNA) and the putative protease Csx29 and catalyzes crRNA-guided RNA cleavage. We report cryo-electron microscopy structures of the Cas7-11-crRNA-Csx29 complex with and without target RNA (tgRNA), and demonstrate that tgRNA binding induces conformational changes in Csx29. Biochemical experiments revealed tgRNA-dependent cleavage of the accessory protein Csx30 by Csx29. Reconstitution of the system in bacteria showed that Csx30 cleavage yields toxic protein fragments that cause growth arrest, which is regulated by Csx31. Csx30 binds Csx31 and the associated sigma factor RpoE (RNA polymerase, extracytoplasmic E), suggesting that Csx30-mediated RpoE inhibition modulates the cellular response to infection. We engineered the Cas7-11-Csx29-Csx30 system for programmable RNA sensing in mammalian cells. Overall, the Cas7-11-Csx29 effector is an RNA-dependent nuclease-protease.

PubMed: 36423304
DOI: 10.1126/science.add7347

実験手法

ELECTRON MICROSCOPY (2.84 Å)