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Structure paper

TitleRNA-activated protein cleavage with a CRISPR-associated endopeptidase.
Journal, issue, pagesScience, Vol. 378, Issue 6622, Page 874-881, Year 2022
Publish dateNov 25, 2022
AuthorsJonathan Strecker / F Esra Demircioglu / David Li / Guilhem Faure / Max E Wilkinson / Jonathan S Gootenberg / Omar O Abudayyeh / Hiroshi Nishimasu / Rhiannon K Macrae / Feng Zhang /
PubMed AbstractIn 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 ...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.
External linksScience / PubMed:36423276 / PubMed Central
MethodsEM (single particle)
Resolution2.53 - 3.15 Å
Structure data

28064

8eex

EMDB-28064: Cas7-11 in complex with Csx29, focus refined on Cas7-11
PDB-8eex: Cas7-11 in complex with Csx29
Method: EM (single particle) / Resolution: 2.95 Å

28065

8eey

EMDB-28065, PDB-8eey:
Cas7-11 in complex with DR-mismatched target RNA, Csx29 and Csx30
Method: EM (single particle) / Resolution: 2.53 Å

EMDB-28070: Cas7-11 in complex with Csx29, focus refined on Csx29
Method: EM (single particle) / Resolution: 3.15 Å

EMDB-28071: Cas7-11 in complex with DR-mismatched target RNA, Csx29 and Csx30, focus refined on Cas7-11 INS domain
Method: EM (single particle) / Resolution: 2.82 Å

EMDB-28072: Cas7-11 in complex with DR-mismatched target RNA, Csx29 and Csx30, focus refined on Csx29
Method: EM (single particle) / Resolution: 2.63 Å

EMDB-28073: Cas7-11 in complex with DR-mismatched target RNA, Csx29 and Csx30, focus refined on Csx29 CHAT domain and Csx30
Method: EM (single particle) / Resolution: 2.72 Å

Chemicals

ChemComp-ZN:
Unknown entry

Source
  • desulfonema ishimotonii (bacteria)
  • synthetic construct (others)
KeywordsRNA BINDING PROTEIN/RNA / CRISPR / endonuclease / endopeptidase / RNA BINDING PROTEIN-RNA complex

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