8Z9C

Cryo-EM structure of NTR-bound type VII CRISPR-Cas complex at substrate-engaged state I

8Z9C の概要

• エントリーDOI: 10.2210/pdb8z9c/pdb
• EMDBエントリー: 39859
• 分子名称: Protein structure, RNA (48-MER), RNA (41-MER), ... (6 entities in total)
• 機能のキーワード: protein structure, antiviral protein
• 由来する生物種: metagenome; 詳細
• タンパク質・核酸の鎖数: 14
• 化学式量合計: 501921.83

構造登録者

Zhang, H.,Deng, Z.,Li, X. (登録日: 2024-04-23, 公開日: 2024-08-21, 最終更新日: 2024-11-27)

主引用文献

Yang, J.,Li, X.,He, Q.,Wang, X.,Tang, J.,Wang, T.,Zhang, Y.,Yu, F.,Zhang, S.,Liu, Z.,Zhang, L.,Liao, F.,Yin, H.,Zhao, H.,Deng, Z.,Zhang, H.
Structural basis for the activity of the type VII CRISPR-Cas system.
Nature, 633:465-472, 2024

PubMed Abstract: The newly identified type VII CRISPR-Cas candidate system uses a CRISPR RNA-guided ribonucleoprotein complex formed by Cas5 and Cas7 proteins to target RNA. However, the RNA cleavage is executed by a dedicated Cas14 nuclease, which is distinct from the effector nucleases of the other CRISPR-Cas systems. Here we report seven cryo-electron microscopy structures of the Cas14-bound interference complex at different functional states. Cas14, a tetrameric protein in solution, is recruited to the Cas5-Cas7 complex in a target RNA-dependent manner. The N-terminal catalytic domain of Cas14 binds a stretch of the substrate RNA for cleavage, whereas the C-terminal domain is primarily responsible for tethering Cas14 to the Cas5-Cas7 complex. The biochemical cleavage assays corroborate the captured functional conformations, revealing that Cas14 binds to different sites on the Cas5-Cas7 complex to execute individual cleavage events. Notably, a plugged-in arginine of Cas7 sandwiched by a C-shaped clamp of C-terminal domain precisely modulates Cas14 binding. More interestingly, target RNA cleavage is altered by a complementary protospacer flanking sequence at the 5' end, but not at the 3' end. Altogether, our study elucidates critical molecular details underlying the assembly of the interference complex and substrate cleavage in the type VII CRISPR-Cas system, which may help rational engineering of the type VII CRISPR-Cas system for biotechnological applications.

PubMed: 39143216
DOI: 10.1038/s41586-024-07815-0

実験手法

ELECTRON MICROSCOPY (3.01 Å)