Characterization of molecular mechanisms controlling fabAB transcription in Pseudomonas aeruginosa

The FabAB pathway is one of the unsaturated fatty acid (UFA) synthesis pathways for Pseudomonas aeruginosa. It was previously noted that this operon was upregulated in biofilms and repressed by exogenous UFAs. Deletion of a 30 nt fabA upstream sequence, which is conserved in P. aeruginosa, P. putida...

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Bibliographic Details
Published in:	PloS one 2012-10, Vol.7 (10), p.e45646
Main Authors:	Schweizer, Herbert P, Choi, Kyoung-Hee
Format:	Article
Language:	eng
Subjects:	3-Oxoacyl-(Acyl-Carrier-Protein) Synthase - genetics 3-Oxoacyl-(Acyl-Carrier-Protein) Synthase - metabolism 5' Untranslated Regions - genetics Amino Acid Sequence Bacterial Proteins - genetics Bacterial Proteins - metabolism Base Sequence Biofilms Biology Biosynthesis Chromosomes Cloning Conserved sequence Deoxyribonucleic acid DNA DNA Transposable Elements - genetics E coli Enzymes Escherichia coli Fatty Acid Synthase, Type II - genetics Fatty Acid Synthase, Type II - metabolism Fatty acids Fatty Acids, Unsaturated - metabolism Gene expression Gene Expression Regulation, Bacterial Gene regulation Genes Genetic engineering Genetic transcription Glycerol Hydro-Lyases - genetics Hydro-Lyases - metabolism Identification methods Medicine Metabolism Molecular modelling Molecular Sequence Data Mutagenesis Mutagenesis, Insertional Nucleic Acid Conformation Operon Plasmids Promoter Regions, Genetic - genetics Protein binding Proteins Pseudomonas aeruginosa Pseudomonas aeruginosa - genetics Pseudomonas aeruginosa - metabolism Pseudomonas putida - genetics Pseudomonas putida - metabolism Pseudomonas syringae - genetics Pseudomonas syringae - metabolism Random mutagenesis Regulatory mechanisms (biology) Regulatory Sequences, Nucleic Acid - genetics Reverse Transcriptase Polymerase Chain Reaction Ribonucleic acid RNA RNA, Bacterial - chemistry RNA, Bacterial - genetics RNA, Bacterial - metabolism Trans-Activators - genetics Trans-Activators - metabolism Transcription Transcription (Genetics) Transcription, Genetic - genetics
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Summary:	The FabAB pathway is one of the unsaturated fatty acid (UFA) synthesis pathways for Pseudomonas aeruginosa. It was previously noted that this operon was upregulated in biofilms and repressed by exogenous UFAs. Deletion of a 30 nt fabA upstream sequence, which is conserved in P. aeruginosa, P. putida, and P. syringae, led to a significant decrease in fabA transcription, suggesting positive regulation by an unknown positive regulatory mechanism. Here, genetic and biochemical approaches were employed to identify a potential fabAB activator. Deletion of candidate genes such as PA1611 or PA1627 was performed to determine if any of these gene products act as a fabAB activator. However, none of these genes were involved in the regulation of fabAB transcription. Use of mariner-based random mutagenesis to screen for fabA activator(s) showed that several genes encoding unknown functions, rpoN and DesA may be involved in fabA regulation, but probably via indirect mechanisms. Biochemical attempts performed did fail to isolate an activator of fabAB operon. The data suggest that fabA expression might not be regulated by protein-binding, but by a distinct mechanism such as a regulatory RNA-based mechanism.
ISSN:	1932-6203 1932-6203