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TitleProtein-templated synthesis of dinucleotide repeat DNA by an antiphage reverse transcriptase.
Journal, issue, pagesScience, Page eaed1656, Year 2026
Publish dateApr 16, 2026
AuthorsPujuan Deng / Hyunbin Lee / Carlo Armijo / Haoqing Wang / Alex Gao /
PubMed AbstractDefense-associated reverse transcriptases (DRTs) are widespread bacterial anti-phage systems that use unconventional mechanisms of polynucleotide synthesis. We show that DRT3, which comprises two ...Defense-associated reverse transcriptases (DRTs) are widespread bacterial anti-phage systems that use unconventional mechanisms of polynucleotide synthesis. We show that DRT3, which comprises two distinct RTs (Drt3a and Drt3b) and a noncoding RNA (ncRNA), synthesizes alternating poly(GT/AC) double-stranded DNA. Cryo-electron microscopy structures at 2.6 Å resolution reveal a D3-symmetric 6:6:6 complex of Drt3a, Drt3b, and ncRNA. Drt3a produces the poly(GT) strand using a conserved ACACAC template within the ncRNA. Notably, Drt3b synthesizes a complementary, protein-primed poly(AC) strand in the complete absence of a nucleic acid template, using conserved active site residues specific to Drt3b to enforce precise base alternation. These findings expand the functional landscape of nucleic acid polymerases, revealing a protein-templated mechanism for sequence-specific DNA synthesis.
External linksScience / PubMed:41990131
MethodsEM (single particle)
Resolution2.49 - 3.02 Å
Structure data

73864

9z6y

EMDB-73864: Cryo-EM hexamer map of the elongating EcDRT3 complex
PDB-9z6y: Structure of the elongating EcDRT3 reverse transcriptase in complex with its non-coding RNA
Method: EM (single particle) / Resolution: 2.6 Å

73865

9z6z

EMDB-73865: Cryo-EM hexamer map of the resting EcDRT3 complex
PDB-9z6z: Structure of the resting EcDRT3 reverse transcriptase in complex with its non-coding RNA
Method: EM (single particle) / Resolution: 2.6 Å

EMDB-75821: Cryo-EM protomer map of the elongating EcDRT3 complex
Method: EM (single particle) / Resolution: 2.49 Å

EMDB-75826: Cryo-EM map of Drt3a and ncRNA in the elongating EcDRT3 complex
Method: EM (single particle) / Resolution: 2.74 Å

EMDB-75828: Cryo-EM protomer map of the resting EcDRT3 complex
Method: EM (single particle) / Resolution: 2.6 Å

EMDB-75830: Cryo-EM map of Drt3a and ncRNA in the resting EcDRT3 complex
Method: EM (single particle) / Resolution: 2.98 Å

EMDB-75981: Cryo-EM map of the EcDRT3 hexameric complex with ddATP and dCTP.
Method: EM (single particle) / Resolution: 2.66 Å

EMDB-75982: Cryo-EM protomer map of the EcDRT3 complex with ddATP and dCTP.
Method: EM (single particle) / Resolution: 2.69 Å

EMDB-75983: Map of the Drt3a-ncRNA subcomplex in the EcDRT3 complex with ddATP and dCTP
Method: EM (single particle) / Resolution: 2.79 Å

EMDB-76001: Composite cryo-EM map of the elongating EcDRT3 complex
Method: EM (single particle) / Resolution: 2.6 Å

EMDB-76002: Composite cryo-EM map of the resting EcDRT3 complex
Method: EM (single particle) / Resolution: 3.02 Å

Chemicals

ChemComp-MG:
Unknown entry

ChemComp-POP:
PYROPHOSPHATE 2-

ChemComp-HOH:
WATER

Source
  • escherichia coli (E. coli)
KeywordsIMMUNE SYSTEM / DRT3-ncRNA complex / reverse transcriptase / Anti-phage complex

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