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TitleCryo-EM structure and protease activity of the type III-E CRISPR-Cas effector.
Journal, issue, pagesNat Microbiol, Vol. 8, Issue 3, Page 522-532, Year 2023
Publish dateJan 26, 2023
AuthorsYangao Huo / Hongshen Zhao / Qinghua Dong / Tao Jiang /
PubMed AbstractThe 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 ...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.
External linksNat Microbiol / PubMed:36702942
MethodsEM (single particle)
Resolution3.14 - 3.64 Å
Structure data

33955

7yn9

EMDB-33955, PDB-7yn9:
Cryo-EM structure of Cas7-11-crRNA binary complex
Method: EM (single particle) / Resolution: 3.53 Å

33956

7yna

EMDB-33956, PDB-7yna:
Cryo-EM structure of Cas7-11-crRNA bound to target RNA-1
Method: EM (single particle) / Resolution: 3.64 Å

33957

7ynb

EMDB-33957, PDB-7ynb:
Cryo-EM structure of Cas7-11-crRNA bound to target RNA-2
Method: EM (single particle) / Resolution: 3.46 Å

33958

7ync

EMDB-33958, PDB-7ync:
Cryo-EM structure of Cas7-11-crRNA bound to target RNA-3
Method: EM (single particle) / Resolution: 3.14 Å

33959

7ynd

EMDB-33959, PDB-7ynd:
Cryo-EM structure of Cas7-11-crRNA-Csx29 ternary complex
Method: EM (single particle) / Resolution: 3.29 Å

Source
  • desulfonema ishimotonii (bacteria)
KeywordsRNA BINDING PROTEIN / Cas7-11 / crRNA / CRISPR-Cas / Csx29

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