Novel targets of the CbrAB/Crc carbon catabolite control system revealed by transcript abundance in Pseudomonas aeruginosa

• Published in: PloS one 2012-10, Vol.7 (10), p.e44637-e44637
• Main Authors: Sonnleitner, Elisabeth, Valentini, Martina, Wenner, Nicolas, Haichar, Feth el Zahar, Haas, Dieter, Lapouge, Karine
• Format: Article
• Language: English
• Subjects: Acids; Amino acids; Analysis; Binding sites; Biofilms; Biology; Branched chain amino acids; Carbon; Carbon - metabolism; Carbon sources; Catabolism; Catabolite repression; Chain branching; Chronic infection; Control systems; Cystic fibrosis; Esterase; Gene expression; Genes, Bacterial; Laboratories; Life Sciences; Lungs; Medicine; Metabolism; Mutation; Nitrogen compounds; Opportunist infection; Proteins; Pseudomonas aeruginosa; Pseudomonas aeruginosa - genetics; Pseudomonas aeruginosa - metabolism; Pseudomonas putida; Ribonucleic acid; RNA; RNA, Messenger - genetics; Signal transduction; Sputum; Substrates; Target recognition; Transcription; Transcription (Genetics)
• ISSN: 1932-6203; 1932-6203
• DOI: 10.1371/journal.pone.0044637

The opportunistic human pathogen Pseudomonas aeruginosa is able to utilize a wide range of carbon and nitrogen compounds, allowing it to grow in vastly different environments. The uptake and catabolism of growth substrates are organized hierarchically by a mechanism termed catabolite repression control (Crc) whereby the Crc protein establishes translational repression of target mRNAs at CA (catabolite activity) motifs present in target mRNAs near ribosome binding sites. Poor carbon sources lead to activation of the CbrAB two-component system, which induces transcription of the small RNA (sRNA) CrcZ. This sRNA relieves Crc-mediated repression of target mRNAs. In this study, we have identified novel targets of the CbrAB/Crc system in P. aeruginosa using transcriptome analysis in combination with a search for CA motifs. We characterized four target genes involved in the uptake and utilization of less preferred carbon sources: estA (secreted esterase), acsA (acetyl-CoA synthetase), bkdR (regulator of branched-chain amino acid catabolism) and aroP2 (aromatic amino acid uptake protein). Evidence for regulation by CbrAB, CrcZ and Crc was obtained in vivo using appropriate reporter fusions, in which mutation of the CA motif resulted in loss of catabolite repression. CbrB and CrcZ were important for growth of P. aeruginosa in cystic fibrosis (CF) sputum medium, suggesting that the CbrAB/Crc system may act as an important regulator during chronic infection of the CF lung.