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Identification of the active mechanism of aminoglycoside entry in V. cholerae through characterization of sRNA ctrR, regulating carbohydrate utilization and transport

The possible active entry of aminoglycosides in bacterial cells has been debated since the development of this antibiotic family. Here we report the identification of their active transport mechanism in Vibrio species. We combined genome-wide transcriptional analysis and fitness screens to identify...

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Autores principales: Pierlé, Sebastian A., Lang, Manon, López-Igual, Rocío, Krin, Evelyne, Fourmy, Dominique, Kennedy, Sean P., Val, Marie-Eve, Baharoglu, Zeynep, Mazel, Didier
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Cold Spring Harbor Laboratory 2023
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10370196/
https://www.ncbi.nlm.nih.gov/pubmed/37502966
http://dx.doi.org/10.1101/2023.07.19.549712
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author Pierlé, Sebastian A.
Lang, Manon
López-Igual, Rocío
Krin, Evelyne
Fourmy, Dominique
Kennedy, Sean P.
Val, Marie-Eve
Baharoglu, Zeynep
Mazel, Didier
author_facet Pierlé, Sebastian A.
Lang, Manon
López-Igual, Rocío
Krin, Evelyne
Fourmy, Dominique
Kennedy, Sean P.
Val, Marie-Eve
Baharoglu, Zeynep
Mazel, Didier
author_sort Pierlé, Sebastian A.
collection PubMed
description The possible active entry of aminoglycosides in bacterial cells has been debated since the development of this antibiotic family. Here we report the identification of their active transport mechanism in Vibrio species. We combined genome-wide transcriptional analysis and fitness screens to identify alterations driven by treatment of V. cholerae with sub-minimum inhibitory concentrations (sub-MIC) of the aminoglycoside tobramycin. RNA-seq data showed downregulation of the small non-coding RNA ncRNA586 during such treatment, while Tn-seq revealed that inactivation of this sRNA was associated with improved fitness in the presence of tobramycin. This sRNA is located near sugar transport genes and previous work on a homologous region in Vibrio tasmaniensis suggested that this sRNA stabilizes gene transcripts for carbohydrate transport and utilization, as well as phage receptors. The role for ncRNA586, hereafter named ctrR, in the transport of both carbohydrates and aminoglycosides, was further investigated. Flow cytometry on cells treated with a fluorescent aminoglycoside confirmed the role of ctrR and of carbohydrate transporters in differential aminoglycoside entry. Despite sequence diversity, ctrR showed functional conservation across the Vibrionales. This system in directly modulated by carbon sources, suggesting regulation by carbon catabolite repression, a widely conserved mechanism in Gram-negative bacteria, priming future research on aminoglycoside uptake by sugar transporters in other bacterial species.
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spelling pubmed-103701962023-07-27 Identification of the active mechanism of aminoglycoside entry in V. cholerae through characterization of sRNA ctrR, regulating carbohydrate utilization and transport Pierlé, Sebastian A. Lang, Manon López-Igual, Rocío Krin, Evelyne Fourmy, Dominique Kennedy, Sean P. Val, Marie-Eve Baharoglu, Zeynep Mazel, Didier bioRxiv Article The possible active entry of aminoglycosides in bacterial cells has been debated since the development of this antibiotic family. Here we report the identification of their active transport mechanism in Vibrio species. We combined genome-wide transcriptional analysis and fitness screens to identify alterations driven by treatment of V. cholerae with sub-minimum inhibitory concentrations (sub-MIC) of the aminoglycoside tobramycin. RNA-seq data showed downregulation of the small non-coding RNA ncRNA586 during such treatment, while Tn-seq revealed that inactivation of this sRNA was associated with improved fitness in the presence of tobramycin. This sRNA is located near sugar transport genes and previous work on a homologous region in Vibrio tasmaniensis suggested that this sRNA stabilizes gene transcripts for carbohydrate transport and utilization, as well as phage receptors. The role for ncRNA586, hereafter named ctrR, in the transport of both carbohydrates and aminoglycosides, was further investigated. Flow cytometry on cells treated with a fluorescent aminoglycoside confirmed the role of ctrR and of carbohydrate transporters in differential aminoglycoside entry. Despite sequence diversity, ctrR showed functional conservation across the Vibrionales. This system in directly modulated by carbon sources, suggesting regulation by carbon catabolite repression, a widely conserved mechanism in Gram-negative bacteria, priming future research on aminoglycoside uptake by sugar transporters in other bacterial species. Cold Spring Harbor Laboratory 2023-07-19 /pmc/articles/PMC10370196/ /pubmed/37502966 http://dx.doi.org/10.1101/2023.07.19.549712 Text en https://creativecommons.org/licenses/by/4.0/This work is licensed under a Creative Commons Attribution 4.0 International License (https://creativecommons.org/licenses/by/4.0/) , which allows reusers to distribute, remix, adapt, and build upon the material in any medium or format, so long as attribution is given to the creator. The license allows for commercial use.
spellingShingle Article
Pierlé, Sebastian A.
Lang, Manon
López-Igual, Rocío
Krin, Evelyne
Fourmy, Dominique
Kennedy, Sean P.
Val, Marie-Eve
Baharoglu, Zeynep
Mazel, Didier
Identification of the active mechanism of aminoglycoside entry in V. cholerae through characterization of sRNA ctrR, regulating carbohydrate utilization and transport
title Identification of the active mechanism of aminoglycoside entry in V. cholerae through characterization of sRNA ctrR, regulating carbohydrate utilization and transport
title_full Identification of the active mechanism of aminoglycoside entry in V. cholerae through characterization of sRNA ctrR, regulating carbohydrate utilization and transport
title_fullStr Identification of the active mechanism of aminoglycoside entry in V. cholerae through characterization of sRNA ctrR, regulating carbohydrate utilization and transport
title_full_unstemmed Identification of the active mechanism of aminoglycoside entry in V. cholerae through characterization of sRNA ctrR, regulating carbohydrate utilization and transport
title_short Identification of the active mechanism of aminoglycoside entry in V. cholerae through characterization of sRNA ctrR, regulating carbohydrate utilization and transport
title_sort identification of the active mechanism of aminoglycoside entry in v. cholerae through characterization of srna ctrr, regulating carbohydrate utilization and transport
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10370196/
https://www.ncbi.nlm.nih.gov/pubmed/37502966
http://dx.doi.org/10.1101/2023.07.19.549712
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