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Structure paper

TitleRibosomal stalk-captured CARF-RelE ribonuclease inhibits translation following CRISPR signaling.
Journal, issue, pagesScience, Vol. 382, Issue 6674, Page 1036-1041, Year 2023
Publish dateNov 30, 2023
AuthorsIrmantas Mogila / Giedre Tamulaitiene / Konstanty Keda / Albertas Timinskas / Audrone Ruksenaite / Giedrius Sasnauskas / Česlovas Venclovas / Virginijus Siksnys / Gintautas Tamulaitis
PubMed AbstractProkaryotic type III CRISPR-Cas antiviral systems employ cyclic oligoadenylate (cA) signaling to activate a diverse range of auxiliary proteins that reinforce the CRISPR-Cas defense. Here we ...Prokaryotic type III CRISPR-Cas antiviral systems employ cyclic oligoadenylate (cA) signaling to activate a diverse range of auxiliary proteins that reinforce the CRISPR-Cas defense. Here we characterize a class of cA-dependent effector proteins named CRISPR-Cas-associated messenger RNA (mRNA) interferase 1 (Cami1) consisting of a CRISPR-associated Rossmann fold sensor domain fused to winged helix-turn-helix and a RelE-family mRNA interferase domain. Upon activation by cyclic tetra-adenylate (cA), Cami1 cleaves mRNA exposed at the ribosomal A-site thereby depleting mRNA and leading to cell growth arrest. The structures of apo-Cami1 and the ribosome-bound Cami1-cA complex delineate the conformational changes that lead to Cami1 activation and the mechanism of Cami1 binding to a bacterial ribosome, revealing unexpected parallels with eukaryotic ribosome-inactivating proteins.
External linksScience / PubMed:38033086
MethodsEM (single particle) / X-ray diffraction
Resolution1.7 - 3.67 Å
Structure data

EMDB-17663: Cami1-ribosome cryoEM-map
Method: EM (single particle) / Resolution: 2.87 Å

EMDB-17664: Cami1-ribosome local refinement map with mask on Cami1 and L12-Cter
Method: EM (single particle) / Resolution: 3.63 Å

EMDB-17665: Cami1-ribosome local refinement map with mask on 30S subunit
Method: EM (single particle) / Resolution: 3.0 Å

EMDB-17666: Cami1-ribosome local refinement map with mask on 50S subunit
Method: EM (single particle) / Resolution: 2.9 Å

17667

8phj

EMDB-17667, PDB-8phj:
cA4-bound Cami1 in complex with 70S ribosome
Method: EM (single particle) / Resolution: 3.67 Å

PDB-8phb:
Crystal structure of apo Cami1
Method: X-RAY DIFFRACTION / Resolution: 1.7 Å

Chemicals

ChemComp-HOH:
WATER / Water

ChemComp-ZN:
Unknown entry

ChemComp-MG:
Unknown entry

Source
  • escherichia coli (E. coli)
  • synthetic construct (others)
  • streptococcus thermophilus (bacteria)
  • allochromatium vinosum (bacteria)
KeywordsHYDROLASE / CRISPR / cyclic oligoadenylate / RNAse / RelE-toxin / RIBOSOME

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