Language

English

Publication Date

11-26-2025

Journal

Nucleic Acids Research

DOI

10.1093/nar/gkaf1227

PMID

41297800

PMCID

PMC12651559

PubMedCentral® Posted Date

11-26-2025

PubMedCentral® Full Text Version

Post-print

Abstract

Double-stranded DNA ends arise from external agents or cellular processes like transcription-replication collisions (TRCs), threatening genome stability. Here, we performed genomic CRISPRi screens to uncover DNA end formation factors in Escherichia coli. We discovered that translation-transcription decoupling causes DNA end formation through a TRC-dependent pathway, which is lethal when DNA end processing by RecBCD is disrupted, but not when recombination is disrupted. We find that TRCs cause replisome stalling followed by "rear-ending" from trailing replisomes which generates free DNA ends, rather than strand breaks. Surprisingly, these DNA ends are resolved through a process we call "replication reset", where the stalled replicore is degraded, without triggering recombination, the DNA damage response, or mutagenesis. This hidden replicore-degradation resets the replication cycle without consequence for the genome. This discovery reveals a novel DNA safeguard mechanism for preserving genome stability when replication is disturbed and challenges the notion that TRCs necessarily cause genome instability in bacteria.

Keywords

DNA Replication, Escherichia coli, Transcription, Genetic, Genomic Instability, DNA, Bacterial, Escherichia coli Proteins, Exodeoxyribonuclease V, DNA Damage

Published Open-Access

yes

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