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PMID: 9029161 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

DNA double-strand breaks caused by replication arrest.

The EMBO journal ·Vol. 16 ·No. 2 ·1997-01-15 ·Pages 430-8

Michel B, Ehrlich SD, Uzest M

Abstract

We report here that DNA double-strand breaks (DSBs) form in Escherichia coli upon arrest of replication forks due to a defect in, or the inhibition of, replicative DNA helicases. The formation of DSBs was assessed by the appearance of linear DNA detected by pulse-field gel electrophoresis. Processing of DSBs by recombination repair or linear DNA degradation was abolished by mutations in recBCD genes. Two E. coli replicative helicases were tested, Rep, which is essential in recBC mutants, and DnaB. The proportion of linear DNA increased up to 50% upon shift of rep recBTS recCTS cells to restrictive temperature. No increase in linear DNA was observed in the absence of replicating chromosomes, indicating that the formation of DSBs in rep strains requires replication. Inhibition of the DnaB helicase either by a strong replication terminator or by a dnaBTS mutation led to the formation of linear DNA, showing that blocked replication forks are prone to DSB formation. In wild-type E. coli, linear DNA was detected in the absence of RecBC or of both RecA and RecD. This reveals the existence of a significant amount of spontaneous DSBs. We propose that some of them may also result from the impairment of replication fork progression.

MeSH Terms
Bacterial Proteins DNA/chemistry DNA Damage DNA Helicases/antagonists & inhibitors DNA Repair DNA Replication DNA, Bacterial/physiology DnaB Helicases Escherichia coli Escherichia coli Proteins Exodeoxyribonuclease V Exodeoxyribonucleases/genetics,metabolism Mutagenesis Rec A Recombinases/genetics,metabolism Temperature
Chemicals
Bacterial Proteins DNA, Bacterial Escherichia coli Proteins DNA Rec A Recombinases Exodeoxyribonucleases Exodeoxyribonuclease V exodeoxyribonuclease V, E coli DNA Helicases DnaB Helicases
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Michel B
Laboratoire de Génétique Microbienne, Institut National de la Recherche Agronomique, Jouy en Josas, France.
Ehrlich S D
Uzest M
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Article Info
Journal
The EMBO journal
Abbr.
EMBO J
ISSN
0261-4189
DOI
10.1093/emboj/16.2.430
Published
1997-01-15
Pages
430-8
Language
English
Region
England
NLM ID
8208664
PMCID
PMC1169647
Subset
IM
External Links
PubMed source DOI: 10.1093/emboj/16.2.430 PMC Full Text View journal details
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