Single-molecule live-cell imaging reveals RecB-dependent function of DNA polymerase IV in double strand break repair.
Ciprofloxacin
/ pharmacology
DNA Breaks, Double-Stranded
/ drug effects
DNA Damage
/ drug effects
DNA Polymerase beta
/ genetics
DNA Repair
/ genetics
DNA Replication
/ genetics
DNA-Binding Proteins
/ genetics
Escherichia coli
/ genetics
Escherichia coli Proteins
/ genetics
Exodeoxyribonuclease V
/ genetics
Rec A Recombinases
/ genetics
Single Molecule Imaging
Journal
Nucleic acids research
ISSN: 1362-4962
Titre abrégé: Nucleic Acids Res
Pays: England
ID NLM: 0411011
Informations de publication
Date de publication:
04 09 2020
04 09 2020
Historique:
accepted:
16
07
2020
revised:
30
06
2020
received:
16
11
2019
pubmed:
21
7
2020
medline:
21
10
2020
entrez:
21
7
2020
Statut:
ppublish
Résumé
Several functions have been proposed for the Escherichia coli DNA polymerase IV (pol IV). Although much research has focused on a potential role for pol IV in assisting pol III replisomes in the bypass of lesions, pol IV is rarely found at the replication fork in vivo. Pol IV is expressed at increased levels in E. coli cells exposed to exogenous DNA damaging agents, including many commonly used antibiotics. Here we present live-cell single-molecule microscopy measurements indicating that double-strand breaks induced by antibiotics strongly stimulate pol IV activity. Exposure to the antibiotics ciprofloxacin and trimethoprim leads to the formation of double strand breaks in E. coli cells. RecA and pol IV foci increase after treatment and exhibit strong colocalization. The induction of the SOS response, the appearance of RecA foci, the appearance of pol IV foci and RecA-pol IV colocalization are all dependent on RecB function. The positioning of pol IV foci likely reflects a physical interaction with the RecA* nucleoprotein filaments that has been detected previously in vitro. Our observations provide an in vivo substantiation of a direct role for pol IV in double strand break repair in cells treated with double strand break-inducing antibiotics.
Identifiants
pubmed: 32687193
pii: 5873806
doi: 10.1093/nar/gkaa597
pmc: PMC7470938
doi:
Substances chimiques
DNA-Binding Proteins
0
Escherichia coli Proteins
0
recA protein, E coli
0
Ciprofloxacin
5E8K9I0O4U
Rec A Recombinases
EC 2.7.7.-
DNA Polymerase beta
EC 2.7.7.7
Exodeoxyribonuclease V
EC 3.1.11.5
exodeoxyribonuclease V, E coli
EC 3.1.11.5
Types de publication
Journal Article
Research Support, N.I.H., Extramural
Research Support, N.I.H., Intramural
Research Support, Non-U.S. Gov't
Langues
eng
Sous-ensembles de citation
IM
Pagination
8490-8508Subventions
Organisme : NIGMS NIH HHS
ID : R01 GM032335
Pays : United States
Organisme : NIEHS NIH HHS
ID : R35 ES028343
Pays : United States
Organisme : NIGMS NIH HHS
ID : RM1 GM130450
Pays : United States
Informations de copyright
© The Author(s) 2020. Published by Oxford University Press on behalf of Nucleic Acids Research.
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