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

Kotecka, Karolina and Kawalek, Adam and Modrzejewska-Balcerek, Magdalena and Gawor, Jan and Zuchniewicz, Karolina and Gromadka, Robert and Bartosik, Aneta A (2022) Functional Characterization of TetR-like Transcriptional Regulator PA3973 from Pseudomonas aeruginosa. International Journal of Molecular Sciences, 23 (14584). pp. 1-25. ISSN 1422-0067

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Official URL: https://www.mdpi.com/1422-0067/23/23/14584

Abstract

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 DPA3972-71 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.

Item Type:Article
Uncontrolled Keywords:Pseudomonas aeruginosa; TetR-like transcriptional regulator; PA3973; gene expression regulation; stress response
Subjects:Q Science > QR Microbiology
Divisions:Department of Microbial Biochemistry
ID Code:2195
Deposited By: PhD AA Bartosik
Deposited On:05 Dec 2022 08:31
Last Modified:05 Dec 2022 08:31

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