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High-throughput screening and whole genome sequencing identifies an antimicrobially active inhibitor of Vibrio cholerae

BACKGROUND: Pathogenic serotypes of Vibrio cholerae cause the life-threatening diarrheal disease cholera. The increasing development of bacterial resistances against the known antibiotics necessitates the search for new antimicrobial compounds and targets for this pathogen. RESULTS: A high-throughpu...

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Autores principales: Sergeev, Galina, Roy, Sambit, Jarek, Michael, Zapolskii, Viktor, Kaufmann, Dieter E, Nandy, Ranjan K, Tegge, Werner
Formato: Online Artículo Texto
Lenguaje:English
Publicado: BioMed Central 2014
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3937525/
https://www.ncbi.nlm.nih.gov/pubmed/24568688
http://dx.doi.org/10.1186/1471-2180-14-49
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author Sergeev, Galina
Roy, Sambit
Jarek, Michael
Zapolskii, Viktor
Kaufmann, Dieter E
Nandy, Ranjan K
Tegge, Werner
author_facet Sergeev, Galina
Roy, Sambit
Jarek, Michael
Zapolskii, Viktor
Kaufmann, Dieter E
Nandy, Ranjan K
Tegge, Werner
author_sort Sergeev, Galina
collection PubMed
description BACKGROUND: Pathogenic serotypes of Vibrio cholerae cause the life-threatening diarrheal disease cholera. The increasing development of bacterial resistances against the known antibiotics necessitates the search for new antimicrobial compounds and targets for this pathogen. RESULTS: A high-throughput screening assay with a Vibrio cholerae reporter strain constitutively expressing green fluorescent protein (GFP) was developed and applied in the investigation of the growth inhibitory effect of approximately 28,300 structurally diverse natural compounds and synthetic small molecules. Several compounds with activities in the low micromolar concentration range were identified. The most active structure, designated vz0825, displayed a minimal inhibitory concentration (MIC) of 1.6 μM and a minimal bactericidal concentration (MBC) of 3.2 μM against several strains of V. cholerae and was specific for this pathogen. Mutants with reduced sensitivity against vz0825 were generated and whole genome sequencing of 15 pooled mutants was carried out. Comparison with the genome of the wild type strain identified the gene VC_A0531 (GenBank: AE003853.1) as the major site of single nucleotide polymorphisms in the resistant mutants. VC_A0531 is located on the small chromosome of V. cholerae and encodes the osmosensitive K(+)-channel sensor histidine kinase (KdpD). Nucleotide exchange of the major mutation site in the wild type strain confirmed the sensitive phenotype. CONCLUSION: The reporter strain MO10 pG13 was successfully used for the identification of new antibacterial compounds against V. cholerae. Generation of resistant mutants and whole genome sequencing was carried out to identify the histidine kinase KdpD as a novel antimicrobial target.
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spelling pubmed-39375252014-03-01 High-throughput screening and whole genome sequencing identifies an antimicrobially active inhibitor of Vibrio cholerae Sergeev, Galina Roy, Sambit Jarek, Michael Zapolskii, Viktor Kaufmann, Dieter E Nandy, Ranjan K Tegge, Werner BMC Microbiol Research Article BACKGROUND: Pathogenic serotypes of Vibrio cholerae cause the life-threatening diarrheal disease cholera. The increasing development of bacterial resistances against the known antibiotics necessitates the search for new antimicrobial compounds and targets for this pathogen. RESULTS: A high-throughput screening assay with a Vibrio cholerae reporter strain constitutively expressing green fluorescent protein (GFP) was developed and applied in the investigation of the growth inhibitory effect of approximately 28,300 structurally diverse natural compounds and synthetic small molecules. Several compounds with activities in the low micromolar concentration range were identified. The most active structure, designated vz0825, displayed a minimal inhibitory concentration (MIC) of 1.6 μM and a minimal bactericidal concentration (MBC) of 3.2 μM against several strains of V. cholerae and was specific for this pathogen. Mutants with reduced sensitivity against vz0825 were generated and whole genome sequencing of 15 pooled mutants was carried out. Comparison with the genome of the wild type strain identified the gene VC_A0531 (GenBank: AE003853.1) as the major site of single nucleotide polymorphisms in the resistant mutants. VC_A0531 is located on the small chromosome of V. cholerae and encodes the osmosensitive K(+)-channel sensor histidine kinase (KdpD). Nucleotide exchange of the major mutation site in the wild type strain confirmed the sensitive phenotype. CONCLUSION: The reporter strain MO10 pG13 was successfully used for the identification of new antibacterial compounds against V. cholerae. Generation of resistant mutants and whole genome sequencing was carried out to identify the histidine kinase KdpD as a novel antimicrobial target. BioMed Central 2014-02-26 /pmc/articles/PMC3937525/ /pubmed/24568688 http://dx.doi.org/10.1186/1471-2180-14-49 Text en Copyright © 2014 Sergeev et al.; licensee BioMed Central Ltd. http://creativecommons.org/licenses/by/2.0 This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly credited.
spellingShingle Research Article
Sergeev, Galina
Roy, Sambit
Jarek, Michael
Zapolskii, Viktor
Kaufmann, Dieter E
Nandy, Ranjan K
Tegge, Werner
High-throughput screening and whole genome sequencing identifies an antimicrobially active inhibitor of Vibrio cholerae
title High-throughput screening and whole genome sequencing identifies an antimicrobially active inhibitor of Vibrio cholerae
title_full High-throughput screening and whole genome sequencing identifies an antimicrobially active inhibitor of Vibrio cholerae
title_fullStr High-throughput screening and whole genome sequencing identifies an antimicrobially active inhibitor of Vibrio cholerae
title_full_unstemmed High-throughput screening and whole genome sequencing identifies an antimicrobially active inhibitor of Vibrio cholerae
title_short High-throughput screening and whole genome sequencing identifies an antimicrobially active inhibitor of Vibrio cholerae
title_sort high-throughput screening and whole genome sequencing identifies an antimicrobially active inhibitor of vibrio cholerae
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3937525/
https://www.ncbi.nlm.nih.gov/pubmed/24568688
http://dx.doi.org/10.1186/1471-2180-14-49
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