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TitleSemirandom DNA adducts regulate a filamentous defense-associated reverse transcriptase.
Journal, issue, pagesNat Struct Mol Biol, Vol. 33, Issue 6, Page 953-961, Year 2026
Publish dateJun 10, 2026
AuthorsNolan Neville / Nicole V Johnson / Edwin E Escobar / Chang-Hwa Chiang / Albana Nreca / Sean R Johnson / Nan Dai / Andy Hanneman / Ivan R Corrêa / Jason S McLellan / Robert J Trachman /
PubMed AbstractRetrons and several defense-associated reverse transcriptases (DRTs) synthesize non-genomic DNA for bacteriophage immunity. In some instances, this non-genomic DNA is of undefined, semirandom ...Retrons and several defense-associated reverse transcriptases (DRTs) synthesize non-genomic DNA for bacteriophage immunity. In some instances, this non-genomic DNA is of undefined, semirandom sequence. How undefined DNA sequences impart antiphage defense is not known. Herewe report the cryo-EM structure and functional characterization of the DRT1 antiphage defense system. We show that DRT1 performs template-free, protein-primed DNA synthesis to generate semirandom DNA adducts. DNA synthesis activates the nitrilase domain of DRT1, while DNA adducts drive the assembly of quiescent DRT1 filaments. Filamentous DRT1 is composed of domain-swapped C termini that are entwined, forming pseudoknots between tetrameric stacks. This configuration occludes conserved active-site residues, resulting in a dormant state. Bacteriophage escape mutants identify a T4 single-stranded DNA helicase required for DRT1 activity. Functionally, DRT1 resembles a minimal retron where a single gene produces a reverse transcriptase, effector and non-genomic antitoxin DNA.
External linksNat Struct Mol Biol / PubMed:42271041 / PubMed Central
MethodsEM (single particle)
Resolution2.6 Å
Structure data

72883

9yfd

EMDB-72883, PDB-9yfd:
Defense-associated reverse transcriptase 1 (DRT1) filament
Method: EM (single particle) / Resolution: 2.6 Å

Chemicals

ChemComp-DTP:
2'-DEOXYADENOSINE 5'-TRIPHOSPHATE

ChemComp-MG:
Unknown entry

Source
  • escherichia coli (E. coli)
KeywordsANTIVIRAL PROTEIN / Anti-phage defense / DNA synthesis / Nitrilase domain / RT domain / filament

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