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Structure paper

TitleRNA-triggered protein cleavage and cell growth arrest by the type III-E CRISPR nuclease-protease.
Journal, issue, pagesScience, Vol. 378, Issue 6622, Page 882-889, Year 2022
Publish dateNov 25, 2022
AuthorsKazuki Kato / Sae Okazaki / Cian Schmitt-Ulms / Kaiyi Jiang / Wenyuan Zhou / Junichiro Ishikawa / Yukari Isayama / Shungo Adachi / Tomohiro Nishizawa / Kira S Makarova / Eugene V Koonin / Omar O Abudayyeh / Jonathan S Gootenberg / Hiroshi Nishimasu /
PubMed AbstractThe type III-E CRISPR-Cas7-11 effector binds a CRISPR RNA (crRNA) and the putative protease Csx29 and catalyzes crRNA-guided RNA cleavage. We report cryo-electron microscopy structures of the Cas7-11- ...The type III-E CRISPR-Cas7-11 effector binds a CRISPR RNA (crRNA) and the putative protease Csx29 and catalyzes crRNA-guided RNA cleavage. We report cryo-electron microscopy structures of the Cas7-11-crRNA-Csx29 complex with and without target RNA (tgRNA), and demonstrate that tgRNA binding induces conformational changes in Csx29. Biochemical experiments revealed tgRNA-dependent cleavage of the accessory protein Csx30 by Csx29. Reconstitution of the system in bacteria showed that Csx30 cleavage yields toxic protein fragments that cause growth arrest, which is regulated by Csx31. Csx30 binds Csx31 and the associated sigma factor RpoE (RNA polymerase, extracytoplasmic E), suggesting that Csx30-mediated RpoE inhibition modulates the cellular response to infection. We engineered the Cas7-11-Csx29-Csx30 system for programmable RNA sensing in mammalian cells. Overall, the Cas7-11-Csx29 effector is an RNA-dependent nuclease-protease.
External linksScience / PubMed:36423304 / PubMed Central
MethodsEM (single particle)
Resolution2.49 - 2.84 Å
Structure data

33695

7y9x

EMDB-33695, PDB-7y9x:
Structure of the Cas7-11-Csx29-guide RNA complex
Method: EM (single particle) / Resolution: 2.49 Å

33696

7y9y

EMDB-33696, PDB-7y9y:
Structure of the Cas7-11-Csx29-guide RNA-target RNA (no PFS) complex
Method: EM (single particle) / Resolution: 2.77 Å

34218

8gs2

EMDB-34218, PDB-8gs2:
Structure of the Cas7-11-Csx29-guide RNA-target RNA (non-matching PFS) complex
Method: EM (single particle) / Resolution: 2.84 Å

Chemicals

ChemComp-ZN:
Unknown entry

ChemComp-C5P:
CYTIDINE-5'-MONOPHOSPHATE

ChemComp-AMP:
ADENOSINE MONOPHOSPHATE / AMP*YM

Source
  • desulfonema ishimotonii (bacteria)
KeywordsRNA BINDING PROTEIN/RNA / CRISPR / RNase / RNA BINDING PROTEIN-RNA COMPLEX

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