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Functional Characterization of TetR-like Transcriptional Regulator PA3973 from Pseudomonas aeruginosa

Pseudomonas aeruginosa, a human opportunistic pathogen, is a common cause of nosocomial infections. Its ability to survive under different conditions relies on a complex regulatory network engaging transcriptional regulators controlling metabolic pathways and capabilities to efficiently use the avai...

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Autores principales: Kotecka, Karolina, Kawalek, Adam, Modrzejewska-Balcerek, Magdalena, Gawor, Jan, Zuchniewicz, Karolina, Gromadka, Robert, Bartosik, Aneta Agnieszka
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9736018/
https://www.ncbi.nlm.nih.gov/pubmed/36498910
http://dx.doi.org/10.3390/ijms232314584
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author Kotecka, Karolina
Kawalek, Adam
Modrzejewska-Balcerek, Magdalena
Gawor, Jan
Zuchniewicz, Karolina
Gromadka, Robert
Bartosik, Aneta Agnieszka
author_facet Kotecka, Karolina
Kawalek, Adam
Modrzejewska-Balcerek, Magdalena
Gawor, Jan
Zuchniewicz, Karolina
Gromadka, Robert
Bartosik, Aneta Agnieszka
author_sort Kotecka, Karolina
collection PubMed
description Pseudomonas aeruginosa, a human opportunistic pathogen, is a common cause of nosocomial infections. Its ability to survive under different conditions relies on a complex regulatory network engaging transcriptional regulators controlling metabolic pathways and capabilities to efficiently use the available resources. P. aeruginosa PA3973 encodes an uncharacterized TetR family transcriptional regulator. In this study, we applied a transcriptome profiling (RNA-seq), genome-wide identification of binding sites using ChIP-seq, as well as the phenotype analyses to unravel the biological role of PA3973. Transcriptional profiling of P. aeruginosa PAO1161 overexpressing PA3973 showed changes in the mRNA level of 648 genes. Concomitantly, ChIP-seq analysis identified more than 300 PA3973 binding sites in the P. aeruginosa genome. A 13 bp sequence motif was indicated as the binding site of PA3973. The PA3973 regulon encompasses the PA3972-PA3971 genes encoding a probable acyl-CoA dehydrogenase and a thioesterase. In vitro analysis showed PA3973 binding to PA3973p. Accordingly, the lack of PA3973 triggered increased expression of PA3972 and PA3971. The ∆PA3972-71 PAO1161 strain demonstrated impaired growth in the presence of stress-inducing agents hydroxylamine or hydroxyurea, thus suggesting the role of PA3972-71 in pathogen survival upon stress. Overall our results showed that TetR-type transcriptional regulator PA3973 has multiple binding sites in the P. aeruginosa genome and influences the expression of diverse genes, including PA3972-PA3971, encoding proteins with a proposed role in stress response.
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spelling pubmed-97360182022-12-11 Functional Characterization of TetR-like Transcriptional Regulator PA3973 from Pseudomonas aeruginosa Kotecka, Karolina Kawalek, Adam Modrzejewska-Balcerek, Magdalena Gawor, Jan Zuchniewicz, Karolina Gromadka, Robert Bartosik, Aneta Agnieszka Int J Mol Sci Article Pseudomonas aeruginosa, a human opportunistic pathogen, is a common cause of nosocomial infections. Its ability to survive under different conditions relies on a complex regulatory network engaging transcriptional regulators controlling metabolic pathways and capabilities to efficiently use the available resources. P. aeruginosa PA3973 encodes an uncharacterized TetR family transcriptional regulator. In this study, we applied a transcriptome profiling (RNA-seq), genome-wide identification of binding sites using ChIP-seq, as well as the phenotype analyses to unravel the biological role of PA3973. Transcriptional profiling of P. aeruginosa PAO1161 overexpressing PA3973 showed changes in the mRNA level of 648 genes. Concomitantly, ChIP-seq analysis identified more than 300 PA3973 binding sites in the P. aeruginosa genome. A 13 bp sequence motif was indicated as the binding site of PA3973. The PA3973 regulon encompasses the PA3972-PA3971 genes encoding a probable acyl-CoA dehydrogenase and a thioesterase. In vitro analysis showed PA3973 binding to PA3973p. Accordingly, the lack of PA3973 triggered increased expression of PA3972 and PA3971. The ∆PA3972-71 PAO1161 strain demonstrated impaired growth in the presence of stress-inducing agents hydroxylamine or hydroxyurea, thus suggesting the role of PA3972-71 in pathogen survival upon stress. Overall our results showed that TetR-type transcriptional regulator PA3973 has multiple binding sites in the P. aeruginosa genome and influences the expression of diverse genes, including PA3972-PA3971, encoding proteins with a proposed role in stress response. MDPI 2022-11-23 /pmc/articles/PMC9736018/ /pubmed/36498910 http://dx.doi.org/10.3390/ijms232314584 Text en © 2022 by the authors. https://creativecommons.org/licenses/by/4.0/Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Kotecka, Karolina
Kawalek, Adam
Modrzejewska-Balcerek, Magdalena
Gawor, Jan
Zuchniewicz, Karolina
Gromadka, Robert
Bartosik, Aneta Agnieszka
Functional Characterization of TetR-like Transcriptional Regulator PA3973 from Pseudomonas aeruginosa
title Functional Characterization of TetR-like Transcriptional Regulator PA3973 from Pseudomonas aeruginosa
title_full Functional Characterization of TetR-like Transcriptional Regulator PA3973 from Pseudomonas aeruginosa
title_fullStr Functional Characterization of TetR-like Transcriptional Regulator PA3973 from Pseudomonas aeruginosa
title_full_unstemmed Functional Characterization of TetR-like Transcriptional Regulator PA3973 from Pseudomonas aeruginosa
title_short Functional Characterization of TetR-like Transcriptional Regulator PA3973 from Pseudomonas aeruginosa
title_sort functional characterization of tetr-like transcriptional regulator pa3973 from pseudomonas aeruginosa
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9736018/
https://www.ncbi.nlm.nih.gov/pubmed/36498910
http://dx.doi.org/10.3390/ijms232314584
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