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TitleProkaryotic Schlafen proteins cleave tRNAs during type III CRISPR immunity.
Journal, issue, pagesNat Commun, Year 2026
Publish dateJul 2, 2026
AuthorsPedro Weickert / Yang Liu / Jonathan Strecker /
PubMed AbstractSchlafen nucleases restrict viral infection in mammals by cleaving self RNAs, however, their function and mechanism in prokaryotic immunity is unknown. Here, we uncover CRISPR-associated Schlafen ...Schlafen nucleases restrict viral infection in mammals by cleaving self RNAs, however, their function and mechanism in prokaryotic immunity is unknown. Here, we uncover CRISPR-associated Schlafen (Cash) proteins containing a Schlafen domain fused to Csx15, an uncharacterized member of Rossmann-like nucleotide-binding sensors. Cash is activated by cyclic tetra-adenylate (cA₄) produced during type III CRISPR interference and induces cell toxicity by cleaving tRNAs, primarily in the T-loop. Cryo-electron microscopy structures of Chloroflexi bacterium Cash reveal an inactive dodecamer, the formation of a filament upon cA₄ binding to align catalytic interfaces, and the molecular basis of substrate recognition and cleavage in a tRNA-bound complex. We identify numerous families of prokaryotic Schlafen proteins associated with diverse antiviral defense systems and characterized by unique sensor domains. This work highlights tRNA depletion by Schlafen nucleases as an evolutionary recurring antiviral strategy and reveals mechanistic differences between Cash and human Schlafen members.
External linksNat Commun / PubMed:42393087
MethodsEM (single particle)
Resolution2.45 - 2.86 Å
Structure data

EMDB-73093, PDB-9ylu:
cryoEM structure of CbCash filament with cA4 and tRNA Ile CAU
Method: EM (single particle) / Resolution: 2.65 Å

EMDB-73094, PDB-9ylv:
Cryo-EM structure of apo CbCash dodecamer
Method: EM (single particle) / Resolution: 2.86 Å

EMDB-73096, PDB-9ylx:
Cryo-EM structure of the CbCash filament bound to cA4
Method: EM (single particle) / Resolution: 2.45 Å

Chemicals

ChemComp-MG:
Unknown entry

Source
  • chloroflexia (bacteria)
  • chloroflexi bacterium 13_1_20cm_2_59_7 (bacteria)
  • escherichia coli bl21(de3) (bacteria)
  • synthetic construct (others)
KeywordsRNA BINDING PROTEIN / tRNA endonuclease / cA4 activation / CRISPR associated Schlafen / Csx15

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