Synergistic effect of SOS response and GATC methylome suppression on antibiotic stress survival in Escherichia coli

The suppression of the SOS response has been shown to enhance the in vitro activity of quinolones. Furthermore, Dam-dependent base methylation has an impact on susceptibility to other antimicrobials affecting DNA synthesis. Here, we investigated the interplay between these two processes, alone and i...

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Authors: Díaz Díaz, S., Recacha, E., Pulido, Marina R., Romero Muñoz, María, De Gregorio Iaria, Belén, Docobo Pérez, Fernando, Pascual Hernández, Álvaro, Rodríguez Martínez, José Manuel
Format: article
Status:Versión aceptada para publicación
Publication Date:2023
Country:España
Institution:Universidad de Sevilla (US)
Repository:idUS. Depósito de Investigación de la Universidad de Sevilla
OAI Identifier:oai:idus.us.es:11441/160849
Online Access:https://hdl.handle.net/11441/160849
https://doi.org/10.1128/aac.01392-22
Access Level:Open access
Keyword:Antibiotic stress
Resistance reversion
Quinolones
RecA gene
Dam methylation system
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spelling Synergistic effect of SOS response and GATC methylome suppression on antibiotic stress survival in Escherichia coliDíaz Díaz, S.Recacha, E.Pulido, Marina R.Romero Muñoz, MaríaDe Gregorio Iaria, BelénDocobo Pérez, FernandoPascual Hernández, ÁlvaroRodríguez Martínez, José ManuelAntibiotic stressResistance reversionQuinolonesRecA geneDam methylation systemThe suppression of the SOS response has been shown to enhance the in vitro activity of quinolones. Furthermore, Dam-dependent base methylation has an impact on susceptibility to other antimicrobials affecting DNA synthesis. Here, we investigated the interplay between these two processes, alone and in combination, in terms of antimicrobial activity. A genetic strategy was used employing single- and double-gene mutants for the SOS response (recA gene) and the Dam methylation system (dam gene) in isogenic models of Escherichia coli both susceptible and resistant to quinolones. Regarding the bacteriostatic activity of quinolones, a synergistic sensitization effect was observed when the Dam methylation system and the recA gene were suppressed. In terms of growth, after 24 h in the presence of quinolones, the Δdam ΔrecA double mutant showed no growth or delayed growth compared to the control strain. In bactericidal terms, spot tests showed that the Δdam ΔrecA double mutant was more sensitive than the ΔrecA single mutant (about 10- to 10² -fold) and the wild type (about 10³- to 10^4-fold) in both susceptible and resistant genetic backgrounds. Differences between the wild type and the Δdam ΔrecA double mutant were confirmed by time-kill assays. The suppression of both systems, in a strain with chromosomal mechanisms of quinolone resistance, prevents the evolution of resistance. This genetic and microbiological approach demonstrated the enhanced sensitization of E. coli to quinolones by dual targeting of the recA (SOS response) and Dam methylation system genes, even in a resistant strain model.Plan Nacional de I1D1i 2013–2016Instituto de Salud Carlos III PI17/01501Instituto de Salud Carlos III PI20/00239Subdirección General de Redes y Centros de Investigación Cooperativa, Ministerio de Economía, Industria y Competitividad, Spanish Network for Research in Infectious Diseases (REIPI) RD16/0016/ 0001Subdirección General de Redes y Centros de Investigación Cooperativa, Ministerio de Economía, Industria y Competitividad, Spanish Network for Research in Infectious Diseases (REIPI) REIPI RD16/0016/0009European Development Regional Fund A Way To Achieve Europe, operative program Intelligent Growth 2014–2020PFIS grant from the Instituto de Salud Carlos III (FI18/00086)ESF Investing in Your FutureJuan Rodés grant from the Instituto de Salud Carlos III (JR21/00030)Universidad de Sevilla VIPPI-US fellowshipondo Social Europeo (EJ5-91, Programa Iniciativa de Empleo Juvenil)American Society of MicrobiologyMicrobiologíaCTS210: Resistencia a Antimicrobianos2023info:eu-repo/semantics/articleinfo:eu-repo/semantics/acceptedVersionapplication/pdfapplication/pdfhttps://hdl.handle.net/11441/160849https://doi.org/10.1128/aac.01392-22reponame:idUS. Depósito de Investigación de la Universidad de Sevillainstname:Universidad de Sevilla (US)InglésAntimicrobial Agents and Chemotherapy, 67 (3), 1-12.info:eu-repo/semantics/openAccessoai:idus.us.es:11441/1608492026-06-17T12:51:07Z
dc.title.none.fl_str_mv Synergistic effect of SOS response and GATC methylome suppression on antibiotic stress survival in Escherichia coli
title Synergistic effect of SOS response and GATC methylome suppression on antibiotic stress survival in Escherichia coli
spellingShingle Synergistic effect of SOS response and GATC methylome suppression on antibiotic stress survival in Escherichia coli
Díaz Díaz, S.
Antibiotic stress
Resistance reversion
Quinolones
RecA gene
Dam methylation system
title_short Synergistic effect of SOS response and GATC methylome suppression on antibiotic stress survival in Escherichia coli
title_full Synergistic effect of SOS response and GATC methylome suppression on antibiotic stress survival in Escherichia coli
title_fullStr Synergistic effect of SOS response and GATC methylome suppression on antibiotic stress survival in Escherichia coli
title_full_unstemmed Synergistic effect of SOS response and GATC methylome suppression on antibiotic stress survival in Escherichia coli
title_sort Synergistic effect of SOS response and GATC methylome suppression on antibiotic stress survival in Escherichia coli
dc.creator.none.fl_str_mv Díaz Díaz, S.
Recacha, E.
Pulido, Marina R.
Romero Muñoz, María
De Gregorio Iaria, Belén
Docobo Pérez, Fernando
Pascual Hernández, Álvaro
Rodríguez Martínez, José Manuel
author Díaz Díaz, S.
author_facet Díaz Díaz, S.
Recacha, E.
Pulido, Marina R.
Romero Muñoz, María
De Gregorio Iaria, Belén
Docobo Pérez, Fernando
Pascual Hernández, Álvaro
Rodríguez Martínez, José Manuel
author_role author
author2 Recacha, E.
Pulido, Marina R.
Romero Muñoz, María
De Gregorio Iaria, Belén
Docobo Pérez, Fernando
Pascual Hernández, Álvaro
Rodríguez Martínez, José Manuel
author2_role author
author
author
author
author
author
author
dc.contributor.none.fl_str_mv Microbiología
CTS210: Resistencia a Antimicrobianos
dc.subject.none.fl_str_mv Antibiotic stress
Resistance reversion
Quinolones
RecA gene
Dam methylation system
topic Antibiotic stress
Resistance reversion
Quinolones
RecA gene
Dam methylation system
description The suppression of the SOS response has been shown to enhance the in vitro activity of quinolones. Furthermore, Dam-dependent base methylation has an impact on susceptibility to other antimicrobials affecting DNA synthesis. Here, we investigated the interplay between these two processes, alone and in combination, in terms of antimicrobial activity. A genetic strategy was used employing single- and double-gene mutants for the SOS response (recA gene) and the Dam methylation system (dam gene) in isogenic models of Escherichia coli both susceptible and resistant to quinolones. Regarding the bacteriostatic activity of quinolones, a synergistic sensitization effect was observed when the Dam methylation system and the recA gene were suppressed. In terms of growth, after 24 h in the presence of quinolones, the Δdam ΔrecA double mutant showed no growth or delayed growth compared to the control strain. In bactericidal terms, spot tests showed that the Δdam ΔrecA double mutant was more sensitive than the ΔrecA single mutant (about 10- to 10² -fold) and the wild type (about 10³- to 10^4-fold) in both susceptible and resistant genetic backgrounds. Differences between the wild type and the Δdam ΔrecA double mutant were confirmed by time-kill assays. The suppression of both systems, in a strain with chromosomal mechanisms of quinolone resistance, prevents the evolution of resistance. This genetic and microbiological approach demonstrated the enhanced sensitization of E. coli to quinolones by dual targeting of the recA (SOS response) and Dam methylation system genes, even in a resistant strain model.
publishDate 2023
dc.date.none.fl_str_mv 2023
dc.type.none.fl_str_mv info:eu-repo/semantics/article
info:eu-repo/semantics/acceptedVersion
format article
status_str acceptedVersion
dc.identifier.none.fl_str_mv https://hdl.handle.net/11441/160849
https://doi.org/10.1128/aac.01392-22
url https://hdl.handle.net/11441/160849
https://doi.org/10.1128/aac.01392-22
dc.language.none.fl_str_mv Inglés
language_invalid_str_mv Inglés
dc.relation.none.fl_str_mv Antimicrobial Agents and Chemotherapy, 67 (3), 1-12.
dc.rights.none.fl_str_mv info:eu-repo/semantics/openAccess
eu_rights_str_mv openAccess
dc.format.none.fl_str_mv application/pdf
application/pdf
dc.publisher.none.fl_str_mv American Society of Microbiology
publisher.none.fl_str_mv American Society of Microbiology
dc.source.none.fl_str_mv reponame:idUS. Depósito de Investigación de la Universidad de Sevilla
instname:Universidad de Sevilla (US)
instname_str Universidad de Sevilla (US)
reponame_str idUS. Depósito de Investigación de la Universidad de Sevilla
collection idUS. Depósito de Investigación de la Universidad de Sevilla
repository.name.fl_str_mv
repository.mail.fl_str_mv
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score 15,812429

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