Nitrate-responsive NarX-NarL represses arginine-mediated induction of the Pseudomonas aeruginosa arginine fermentation arcDABC operon

• Published in: Microbiology (Society for General Microbiology) 2008-10, Vol.154 (10), p.3053-3060
• Main Authors: Benkert, Beatrice, Quack, Nicole, Schreiber, Kerstin, Jaensch, Lothar, Jahn, Dieter, Schobert, Max
• Format: Article
• Language: English
• Subjects: Amino Acid Transport Systems - genetics; Amino Acid Transport Systems - metabolism; Anaerobiosis; Antiporters - genetics; Antiporters - metabolism; Arginine - metabolism; Bacterial Proteins - genetics; Bacterial Proteins - metabolism; Bacteriology; Binding Sites; Biological and medical sciences; Blotting, Northern; Fermentation; Fundamental and applied biological sciences. Psychology; Gene Expression Regulation, Bacterial; Genes, Bacterial; Genes, Reporter; Genetics; Microbiology; Mutagenesis, Site-Directed; Nitrates - metabolism; Operon; Promoter Regions, Genetic; Proteome - genetics; Proteome - metabolism; Pseudomonas aeruginosa; Pseudomonas aeruginosa - genetics; Pseudomonas aeruginosa - metabolism; Repressor Proteins
• ISSN: 1350-0872; 1465-2080
• DOI: 10.1099/mic.0.2008/018929-0

1 Institute of Microbiology, Technical University Braunschweig, Spielmannstr. 7, D-38106 Braunschweig, Germany 2 Division of Cell and Immune Biology, Proteome Research Group, Helmholtz Centre for Infection Research, Inhoffenstr. 7, D-38124 Braunschweig, Germany Correspondence Max Schobert m.schobert{at}tu-bs.de Denitrification and arginine fermentation are major parts of the anaerobic metabolism of Pseudomonas aeruginosa , which is important for biofilm formation and infection. The two-component regulatory system NarX-NarL is part of the underlying network and is required for denitrifying growth. All target promoters identified so far are activated by NarL. In this study the effect of NarL on arginine fermentation was investigated using proteome, Northern blot and lacZ reporter gene analyses. NarL-dependent repression of the arcDABC operon was observed and the corresponding NarL-binding site in the arcD promoter region was functionally localized at –60 bp upstream of the transcriptional start site using site-directed promoter mutagenesis and reporter gene fusion experiments. The results clearly show that in the presence of nitrate NarL represses the arginine-dependent activation of the arcDABC operon mediated by ArgR. It does not influence the oxygen-tension-dependent activation via Anr. Thus, the anaerobic energy metabolism of P. aeruginosa is coordinated via NarX-NarL activity. In the presence of nitrate the highly efficient denitrification is preferred over the less attractive arginine fermentation. Abbreviations: MU, Miller units; RuBPS, ruthenium(II)-tris(bathophenanthroline disulfonate) Present address: Institute of Biochemistry and Biotechnology, Technical University Braunschweig, Spielmannstr. 7, D-38106 Braunschweig, Germany.