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TitleDual sensing activates antiviral reverse transcriptase for membrane targeting.
Journal, issue, pagesCell Host Microbe, Year 2026
Publish dateSep 2, 2026
AuthorsYushan Xia / Hua Qi / Xin-Yang Wei / Jun-Tao Zhang / Wenao Hu / Yu-Jun Liu / Xin-Yi Song / Xue Liu / Chongyuan Wang / Ning Jia /
PubMed AbstractThe prokaryotic type 2 defense-associated reverse transcriptase (DRT2) system mediates antiviral defense by catalyzing the rolling-circle reverse transcription of a noncoding RNA (ncRNA) and ...The prokaryotic type 2 defense-associated reverse transcriptase (DRT2) system mediates antiviral defense by catalyzing the rolling-circle reverse transcription of a noncoding RNA (ncRNA) and producing the toxic Neo protein that arrests bacterial growth. However, the mechanisms underlying DRT2 activation and effector function remain unknown. Here, we identified two distinct activation mechanisms: direct binding of a phage-encoded single-stranded DNA-binding protein (SSB or SSAP), and the detection of elevated intracellular dGTP levels induced by the phage-encoded ribonucleotide reductase NrdAB. Upon activation, the produced Neo protein directly targets the bacterial membrane, inducing membrane depolarization and growth arrest. Cryo-electron microscopy (cryo-EM) structures of the DRT2-ncRNA complex in its arrested and dGTP-bound active states provide mechanistic insights into rolling-circle ccDNA synthesis and template jumping. Furthermore, these identified activation mechanisms enable the DRT2 system with an engineered ncRNA template to produce a large-scale, user-defined double-stranded DNA (dsDNA) template in vivo, highlighting its potential in biotechnological applications.
External linksCell Host Microbe / PubMed:42685694
MethodsEM (single particle)
Resolution2.75 - 3.52 Å
Structure data

63146

9lje

EMDB-63146, PDB-9lje:
Cryo-EM structure of the DRT2-ncRNA complex
Method: EM (single particle) / Resolution: 2.75 Å

63147

9ljf

EMDB-63147, PDB-9ljf:
Cryo-EM structure of the dGTP bound DRT2-ncRNA complex
Method: EM (single particle) / Resolution: 3.52 Å

Chemicals

ChemComp-MG:
Unknown entry

ChemComp-DGT:
2'-DEOXYGUANOSINE-5'-TRIPHOSPHATE

Source
  • klebsiella pneumoniae (bacteria)
KeywordsANTIVIRAL PROTEIN/RNA/DNA / RNA BINDING PROTEIN-RNA-DNA COMPLEX / ANTIVIRAL PROTEIN-RNA-DNA complex

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