Pseudomonas aeruginosa LysR PA4203 regulator NmoR acts as a repressor of the PA4202 nmoA gene, encoding a nitronate monooxygenase

Ken Vercammen; Qing Wei; Daniel Charlier; Andreas Dötsch; Susanne Haüssler; Sebastian Schulz; Francesca Salvi; Giovanni Gadda; Jim Spain; Morten Levin Rybtke; Tim Tolker-Nielsen; Jozef Dingemans; Lumeng Ye; Pierre Cornelis

doi:10.1128/JB.01991-14

Pseudomonas aeruginosa LysR PA4203 regulator NmoR acts as a repressor of the PA4202 nmoA gene, encoding a nitronate monooxygenase

Ken Vercammen, Qing Wei, Daniel Charlier, Andreas Dötsch, Susanne Haüssler, Sebastian Schulz, Francesca Salvi, Giovanni Gadda, Jim Spain, Morten Levin Rybtke, Tim Tolker-Nielsen, Jozef Dingemans, Lumeng Ye, Pierre Cornelis

3 Citations (Scopus)

Abstract

The PA4203 gene encodes a LysR regulator and lies between the ppgL gene (PA4204), which encodes a periplasmic gluconolactonase, and, in the opposite orientation, the PA4202 (nmoA) gene, coding for a nitronate monooxygenase, and ddlA (PA4201), encoding a d-alanine alanine ligase. The intergenic regions between PA4203 and ppgL and between PA4203 and nmoA are very short (79 and 107 nucleotides, respectively). Here we show that PA4203 (nmoR) represses its own transcription and the expression of nmoA. A chromatin immunoprecipitation analysis showed the presence of a single NmoR binding site between nmoA and nmoR, which was confirmed by electrophoretic mobility shift assays (EMSAs) with the purified NmoR protein. Despite this observation, a transcriptome analysis revealed more genes to be affected in an nmoR mutant, including genes known to be part of the MexT LysR activator regulon. The PA1225 gene, encoding a quinone oxidoreductase, was the most highly upregulated gene in the nmoR deletion mutant, independently of MexT. Finally, deletion of the nmoA gene resulted in an increased sensitivity of the cells to 3-nitropropionic acid (3-NPA), confirming the role of the nitronate monooxygenase protein in the detoxification of nitronate.

Original language	English
Journal	Journal of Bacteriology
Volume	197
Issue number	6
Pages (from-to)	1026-39
Number of pages	14
ISSN	0021-9193
DOIs	https://doi.org/10.1128/JB.01991-14
Publication status	Published - Mar 2015

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Vercammen, K., Wei, Q., Charlier, D., Dötsch, A., Haüssler, S., Schulz, S., Salvi, F., Gadda, G., Spain, J., Rybtke, M. L., Tolker-Nielsen, T., Dingemans, J., Ye, L., & Cornelis, P. (2015). Pseudomonas aeruginosa LysR PA4203 regulator NmoR acts as a repressor of the PA4202 nmoA gene, encoding a nitronate monooxygenase. Journal of Bacteriology, 197(6), 1026-39. https://doi.org/10.1128/JB.01991-14

Vercammen, K, Wei, Q, Charlier, D, Dötsch, A, Haüssler, S, Schulz, S, Salvi, F, Gadda, G, Spain, J, Rybtke, ML , Tolker-Nielsen, T, Dingemans, J, Ye, L & Cornelis, P 2015, 'Pseudomonas aeruginosa LysR PA4203 regulator NmoR acts as a repressor of the PA4202 nmoA gene, encoding a nitronate monooxygenase', Journal of Bacteriology, vol. 197, no. 6, pp. 1026-39. https://doi.org/10.1128/JB.01991-14

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title = "Pseudomonas aeruginosa LysR PA4203 regulator NmoR acts as a repressor of the PA4202 nmoA gene, encoding a nitronate monooxygenase",

abstract = "The PA4203 gene encodes a LysR regulator and lies between the ppgL gene (PA4204), which encodes a periplasmic gluconolactonase, and, in the opposite orientation, the PA4202 (nmoA) gene, coding for a nitronate monooxygenase, and ddlA (PA4201), encoding a d-alanine alanine ligase. The intergenic regions between PA4203 and ppgL and between PA4203 and nmoA are very short (79 and 107 nucleotides, respectively). Here we show that PA4203 (nmoR) represses its own transcription and the expression of nmoA. A chromatin immunoprecipitation analysis showed the presence of a single NmoR binding site between nmoA and nmoR, which was confirmed by electrophoretic mobility shift assays (EMSAs) with the purified NmoR protein. Despite this observation, a transcriptome analysis revealed more genes to be affected in an nmoR mutant, including genes known to be part of the MexT LysR activator regulon. The PA1225 gene, encoding a quinone oxidoreductase, was the most highly upregulated gene in the nmoR deletion mutant, independently of MexT. Finally, deletion of the nmoA gene resulted in an increased sensitivity of the cells to 3-nitropropionic acid (3-NPA), confirming the role of the nitronate monooxygenase protein in the detoxification of nitronate.",

author = "Ken Vercammen and Qing Wei and Daniel Charlier and Andreas D{\"o}tsch and Susanne Ha{\"u}ssler and Sebastian Schulz and Francesca Salvi and Giovanni Gadda and Jim Spain and Rybtke, {Morten Levin} and Tim Tolker-Nielsen and Jozef Dingemans and Lumeng Ye and Pierre Cornelis",

year = "2015",

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doi = "10.1128/JB.01991-14",

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T1 - Pseudomonas aeruginosa LysR PA4203 regulator NmoR acts as a repressor of the PA4202 nmoA gene, encoding a nitronate monooxygenase

AU - Vercammen, Ken

AU - Wei, Qing

AU - Charlier, Daniel

AU - Dötsch, Andreas

AU - Haüssler, Susanne

AU - Schulz, Sebastian

AU - Salvi, Francesca

AU - Gadda, Giovanni

AU - Spain, Jim

AU - Rybtke, Morten Levin

AU - Tolker-Nielsen, Tim

AU - Dingemans, Jozef

AU - Ye, Lumeng

AU - Cornelis, Pierre

PY - 2015/3

Y1 - 2015/3

N2 - The PA4203 gene encodes a LysR regulator and lies between the ppgL gene (PA4204), which encodes a periplasmic gluconolactonase, and, in the opposite orientation, the PA4202 (nmoA) gene, coding for a nitronate monooxygenase, and ddlA (PA4201), encoding a d-alanine alanine ligase. The intergenic regions between PA4203 and ppgL and between PA4203 and nmoA are very short (79 and 107 nucleotides, respectively). Here we show that PA4203 (nmoR) represses its own transcription and the expression of nmoA. A chromatin immunoprecipitation analysis showed the presence of a single NmoR binding site between nmoA and nmoR, which was confirmed by electrophoretic mobility shift assays (EMSAs) with the purified NmoR protein. Despite this observation, a transcriptome analysis revealed more genes to be affected in an nmoR mutant, including genes known to be part of the MexT LysR activator regulon. The PA1225 gene, encoding a quinone oxidoreductase, was the most highly upregulated gene in the nmoR deletion mutant, independently of MexT. Finally, deletion of the nmoA gene resulted in an increased sensitivity of the cells to 3-nitropropionic acid (3-NPA), confirming the role of the nitronate monooxygenase protein in the detoxification of nitronate.

AB - The PA4203 gene encodes a LysR regulator and lies between the ppgL gene (PA4204), which encodes a periplasmic gluconolactonase, and, in the opposite orientation, the PA4202 (nmoA) gene, coding for a nitronate monooxygenase, and ddlA (PA4201), encoding a d-alanine alanine ligase. The intergenic regions between PA4203 and ppgL and between PA4203 and nmoA are very short (79 and 107 nucleotides, respectively). Here we show that PA4203 (nmoR) represses its own transcription and the expression of nmoA. A chromatin immunoprecipitation analysis showed the presence of a single NmoR binding site between nmoA and nmoR, which was confirmed by electrophoretic mobility shift assays (EMSAs) with the purified NmoR protein. Despite this observation, a transcriptome analysis revealed more genes to be affected in an nmoR mutant, including genes known to be part of the MexT LysR activator regulon. The PA1225 gene, encoding a quinone oxidoreductase, was the most highly upregulated gene in the nmoR deletion mutant, independently of MexT. Finally, deletion of the nmoA gene resulted in an increased sensitivity of the cells to 3-nitropropionic acid (3-NPA), confirming the role of the nitronate monooxygenase protein in the detoxification of nitronate.

U2 - 10.1128/JB.01991-14

DO - 10.1128/JB.01991-14

M3 - Journal article

C2 - 25384477

SN - 0021-9193

VL - 197

SP - 1026

EP - 1039

JO - Journal of Bacteriology

JF - Journal of Bacteriology

IS - 6

ER -

Pseudomonas aeruginosa LysR PA4203 regulator NmoR acts as a repressor of the PA4202 nmoA gene, encoding a nitronate monooxygenase

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