JPOST Repositoryapplication/xmlhttps://storage.jpostdb.org/JPST004145/files/LFQResults__May2025.xlsxhttps://storage.jpostdb.org/JPST004145/files/20250527-S03_01.rawhttps://storage.jpostdb.org/JPST004145/files/20250527-S02_01.rawhttps://storage.jpostdb.org/JPST004145/files/20250527-S01_01.rawhttps://storage.jpostdb.org/JPST004145/files/20250527-S13_01.rawhttps://storage.jpostdb.org/JPST004145/files/20250527-S12_01.rawhttps://storage.jpostdb.org/JPST004145/files/20250527-S11_01.rawhttps://storage.jpostdb.org/JPST004145/files/20250527-S10_01.rawhttps://storage.jpostdb.org/JPST004145/files/20250527-S09_01.rawhttps://storage.jpostdb.org/JPST004145/files/20250527-S08_01.rawhttps://storage.jpostdb.org/JPST004145/files/20250527-S07_01.rawhttps://storage.jpostdb.org/JPST004145/files/20250527-S06_01.rawhttps://storage.jpostdb.org/JPST004145/files/20250527-S04_01.rawhttps://storage.jpostdb.org/JPST004145/files/20250527-S05_01.rawhttps://storage.jpostdb.org/JPST004145/files/20250527-S14_01.rawhttps://storage.jpostdb.org/JPST004145/files/20250527-S15_01.rawprimary200OKProteomicsRobert TrachmanEscherichia Colihttps://repository.jpostdb.org/entry/JPST004145New England BiolabsjPOSTRetrons 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.Semirandom DNA adducts regulate a filamentous defense-associated reverse transcriptase.Neville Nolan N, Johnson Nicole V NV, Escobar Edwin E EE, Chiang Chang-Hwa CH, Nreca Albana A, Johnson Sean R SR, Dai Nan N, Hanneman Andy A, Corrêa Ivan R IR, McLellan Jason S JS, Trachman Robert J RJfalseDRT1 FLAG Pulldown Raw Data from Exploris 480We aim to see what T4 phage and/or E. coli proteins interact with our DRT1 enzyme. We have conducted a pulldown assay using N-terminally MBP-tagged DRT1, both for WT and RT-inactive mutant. Fri Jul 10 00:00:00 BST 2026PXD07003956242271041