7YND

Cryo-EM structure of Cas7-11-crRNA-Csx29 ternary complex

Summary for 7YND

• Entry DOI: 10.2210/pdb7ynd/pdb
• EMDB information: 33959
• Descriptor: CRISPR-associated RAMP family protein, crRNA (38-MER), CHAT domain-containing protein (3 entities in total)
• Functional Keywords: cas7-11, crrna, crispr-cas, csx29, rna binding protein
• Biological source: Desulfonema ishimotonii; More
• Total number of polymer chains: 3
• Total formula weight: 287338.91

Authors

Huo, Y.,Dong, Q.,Zhao, H.,Jiang, T. (deposition date: 2022-07-30, release date: 2023-02-01, Last modification date: 2024-10-16)

Primary citation

Huo, Y.,Zhao, H.,Dong, Q.,Jiang, T.
Cryo-EM structure and protease activity of the type III-E CRISPR-Cas effector.
Nat Microbiol, 8:522-532, 2023

PubMed Abstract: The recently discovered type III-E CRISPR-Cas effector Cas7-11 shows promise when used as an RNA manipulation tool, but its structure and the mechanisms underlying its function remain unclear. Here we present four cryo-EM structures of Desulfonema ishimotonii Cas7-11-crRNA complex in pre-target and target RNA-bound states, and the cryo-EM structure of DiCas7-11-crRNA bound to its accessory protein DiCsx29. These data reveal structural elements for pre-crRNA processing, target RNA cleavage and regulation. Moreover, a 3' seed region of crRNA is involved in regulating RNA cleavage activity of DiCas7-11-crRNA-Csx29. Our analysis also shows that both the minimal mismatch of 4 nt to the 5' handle of crRNA and the minimal matching of the first 12 nt of the spacer by the target RNA are essential for triggering the protease activity of DiCas7-11-crRNA-Csx29 towards DiCsx30. Taken together, we propose that target RNA recognition and cleavage regulate and fine-tune the protease activity of DiCas7-11-crRNA-Csx29, thus preventing auto-immune responses.

PubMed: 36702942
DOI: 10.1038/s41564-022-01316-4

Experimental method

ELECTRON MICROSCOPY (3.29 Å)