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Putrescine biosynthesis and export genes are essential for normal growth of avian pathogenic Escherichia coli

BACKGROUND: Avian pathogenic Escherichia coli (APEC) is the infectious agent of a wide variety of avian diseases, which causes substantial economic losses to the poultry industry worldwide. Polyamines contribute to the optimal synthesis of nucleic acids and proteins in bacteria. The objectives of th...

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Autores principales: Guerra, Priscila R., Herrero-Fresno, Ana, Ladero, Victor, Redruello, Begoña, dos Santos, Teresa Pires, Spiegelhauer, Malene R., Jelsbak, Lotte, Olsen, John Elmerdahl
Formato: Online Artículo Texto
Lenguaje:English
Publicado: BioMed Central 2018
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6307189/
https://www.ncbi.nlm.nih.gov/pubmed/30587122
http://dx.doi.org/10.1186/s12866-018-1355-9
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author Guerra, Priscila R.
Herrero-Fresno, Ana
Ladero, Victor
Redruello, Begoña
dos Santos, Teresa Pires
Spiegelhauer, Malene R.
Jelsbak, Lotte
Olsen, John Elmerdahl
author_facet Guerra, Priscila R.
Herrero-Fresno, Ana
Ladero, Victor
Redruello, Begoña
dos Santos, Teresa Pires
Spiegelhauer, Malene R.
Jelsbak, Lotte
Olsen, John Elmerdahl
author_sort Guerra, Priscila R.
collection PubMed
description BACKGROUND: Avian pathogenic Escherichia coli (APEC) is the infectious agent of a wide variety of avian diseases, which causes substantial economic losses to the poultry industry worldwide. Polyamines contribute to the optimal synthesis of nucleic acids and proteins in bacteria. The objectives of this study were to investigate; i) whether APEC E. coli encodes the same systems for biosynthesis and uptake as described for E. coli K12 and ii) the role of polyamines during in vitro growth of an avian pathogenic E. coli strain (WT-ST117- O83:H4T). RESULTS: Following whole genome sequencing, polyamine biosynthesis and export genes present in E. coli MG1655 (K-12) were found to be identical in WT-ST117. Defined mutants were constructed in putrescine and spermidine biosynthesis pathways (ΔspeB, ΔspeC, ΔspeF, ΔspeB/C and ΔspeD/E), and in polyamines transport systems (ΔpotE, ΔyeeF, ΔpotABCD and ΔpotFGHI). Contrary to what was observed for MG1655, the ΔpotE-ST117 mutant was growth attenuated, regardless of putrescine supplementation. The addition of spermidine or orthinine restored the growth to the level of WT-ST117. Growth attenuation after induction of membrane stress by SDS suggested that PotE is involved in protection against this stress. The ΔspeB/C-ST117 mutant was also growth attenuated in minimal medium. The addition of putrescine or spermidine to the media restored growth rate to the wild type level. The remaining biosynthesis and transport mutants showed a growth similar to that of WT-ST117. Analysis by Ultra-High Performance Liquid Chromatography revealed that the ΔspeB/C mutant was putrescine-deficient, despite that the gene speF, which is also involved in the synthesis of putrescine, was expressed. CONCLUSIONS: Deletion of the putrescine transport system, PotE, or the putrescine biosynthesis pathway genes speB/C affected in vitro growth of APEC (ST117- O83:H4) strain, but not E. coli MG1655, despite the high similarity of the genetic make-up of biosynthesis and transport genes. Therefore, blocking these metabolic reactions may be a suitable way to prevent APEC growth in the host without disturbing the commensal E. coli population. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (10.1186/s12866-018-1355-9) contains supplementary material, which is available to authorized users.
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spelling pubmed-63071892019-01-02 Putrescine biosynthesis and export genes are essential for normal growth of avian pathogenic Escherichia coli Guerra, Priscila R. Herrero-Fresno, Ana Ladero, Victor Redruello, Begoña dos Santos, Teresa Pires Spiegelhauer, Malene R. Jelsbak, Lotte Olsen, John Elmerdahl BMC Microbiol Research Article BACKGROUND: Avian pathogenic Escherichia coli (APEC) is the infectious agent of a wide variety of avian diseases, which causes substantial economic losses to the poultry industry worldwide. Polyamines contribute to the optimal synthesis of nucleic acids and proteins in bacteria. The objectives of this study were to investigate; i) whether APEC E. coli encodes the same systems for biosynthesis and uptake as described for E. coli K12 and ii) the role of polyamines during in vitro growth of an avian pathogenic E. coli strain (WT-ST117- O83:H4T). RESULTS: Following whole genome sequencing, polyamine biosynthesis and export genes present in E. coli MG1655 (K-12) were found to be identical in WT-ST117. Defined mutants were constructed in putrescine and spermidine biosynthesis pathways (ΔspeB, ΔspeC, ΔspeF, ΔspeB/C and ΔspeD/E), and in polyamines transport systems (ΔpotE, ΔyeeF, ΔpotABCD and ΔpotFGHI). Contrary to what was observed for MG1655, the ΔpotE-ST117 mutant was growth attenuated, regardless of putrescine supplementation. The addition of spermidine or orthinine restored the growth to the level of WT-ST117. Growth attenuation after induction of membrane stress by SDS suggested that PotE is involved in protection against this stress. The ΔspeB/C-ST117 mutant was also growth attenuated in minimal medium. The addition of putrescine or spermidine to the media restored growth rate to the wild type level. The remaining biosynthesis and transport mutants showed a growth similar to that of WT-ST117. Analysis by Ultra-High Performance Liquid Chromatography revealed that the ΔspeB/C mutant was putrescine-deficient, despite that the gene speF, which is also involved in the synthesis of putrescine, was expressed. CONCLUSIONS: Deletion of the putrescine transport system, PotE, or the putrescine biosynthesis pathway genes speB/C affected in vitro growth of APEC (ST117- O83:H4) strain, but not E. coli MG1655, despite the high similarity of the genetic make-up of biosynthesis and transport genes. Therefore, blocking these metabolic reactions may be a suitable way to prevent APEC growth in the host without disturbing the commensal E. coli population. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (10.1186/s12866-018-1355-9) contains supplementary material, which is available to authorized users. BioMed Central 2018-12-27 /pmc/articles/PMC6307189/ /pubmed/30587122 http://dx.doi.org/10.1186/s12866-018-1355-9 Text en © The Author(s). 2018 Open AccessThis article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.
spellingShingle Research Article
Guerra, Priscila R.
Herrero-Fresno, Ana
Ladero, Victor
Redruello, Begoña
dos Santos, Teresa Pires
Spiegelhauer, Malene R.
Jelsbak, Lotte
Olsen, John Elmerdahl
Putrescine biosynthesis and export genes are essential for normal growth of avian pathogenic Escherichia coli
title Putrescine biosynthesis and export genes are essential for normal growth of avian pathogenic Escherichia coli
title_full Putrescine biosynthesis and export genes are essential for normal growth of avian pathogenic Escherichia coli
title_fullStr Putrescine biosynthesis and export genes are essential for normal growth of avian pathogenic Escherichia coli
title_full_unstemmed Putrescine biosynthesis and export genes are essential for normal growth of avian pathogenic Escherichia coli
title_short Putrescine biosynthesis and export genes are essential for normal growth of avian pathogenic Escherichia coli
title_sort putrescine biosynthesis and export genes are essential for normal growth of avian pathogenic escherichia coli
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6307189/
https://www.ncbi.nlm.nih.gov/pubmed/30587122
http://dx.doi.org/10.1186/s12866-018-1355-9
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