8EEY

Cas7-11 in complex with DR-mismatched target RNA, Csx29 and Csx30

This is a non-PDB format compatible entry.

Summary for 8EEY

• Entry DOI: 10.2210/pdb8eey/pdb
• EMDB information: 28065
• Descriptor: Csx30, Cas7-11, Csx29, ... (6 entities in total)
• Functional Keywords: crispr, endonuclease, endopeptidase, rna binding protein-rna complex, rna binding protein/rna
• Biological source: Desulfonema ishimotonii; More
• Total number of polymer chains: 5
• Total formula weight: 356172.19

Authors

Demircioglu, F.E.,Wilkinson, M.E.,Strecker, J.,Li, D.,Faure, G.,Macrae, R.K.,Zhang, F. (deposition date: 2022-09-07, release date: 2022-11-16, Last modification date: 2024-06-19)

Primary citation

Strecker, J.,Demircioglu, F.E.,Li, D.,Faure, G.,Wilkinson, M.E.,Gootenberg, J.S.,Abudayyeh, O.O.,Nishimasu, H.,Macrae, R.K.,Zhang, F.
RNA-activated protein cleavage with a CRISPR-associated endopeptidase.
Science, 378:874-881, 2022

PubMed Abstract: In prokaryotes, CRISPR-Cas systems provide adaptive immune responses against foreign genetic elements through RNA-guided nuclease activity. Recently, additional genes with non-nuclease functions have been found in genetic association with CRISPR systems, suggesting that there may be other RNA-guided non-nucleolytic enzymes. One such gene from encodes the TPR-CHAT protease Csx29, which is associated with the CRISPR effector Cas7-11. Here, we demonstrate that this CRISPR-associated protease (CASP) exhibits programmable RNA-activated endopeptidase activity against a sigma factor inhibitor to regulate a transcriptional response. Cryo-electron microscopy of an active and substrate-bound CASP complex reveals an allosteric activation mechanism that reorganizes Csx29 catalytic residues upon target RNA binding. This work reveals an RNA-guided function in nature that can be leveraged for RNA-sensing applications in vitro and in human cells.

PubMed: 36423276
DOI: 10.1126/science.add7450

Experimental method

ELECTRON MICROSCOPY (2.53 Å)