title: In vitro phosphorylation of AlgR, a regulator of mucoidy in Pseudomonas aeruginosa, by a histidine protein kinase and effects of small phospho-donor molecules.
abstract: AlgR is a transcriptional regulator of mucoidy in Pseudomonas aeruginosa, a critical virulence factor expressed in cystic fibrosis. AlgR belongs to the superfamily of bacterial signal transduction systems, and has been shown to bind to the algD promoter, a critical point in the regulation of mucoidy. This protein, like other typical response regulators, contains highly conserved residues known to be critical for the phosphorylation and signal transduction processes. However, a typical second component interacting with AlgR has not been identified. Here we demonstrate that AlgR undergoes phosphorylation in vitro when interacting with the well-characterized histidine protein kinase CheA. These results indicate that AlgR is capable of undergoing phosphorylation typical of other two-component signal transduction systems. Moreover, the phosphotransfer reaction between CheA and AlgR was found to be affected by the presence of carbamoyl phosphate, acetyl phosphate, and salts of phosphoramidic acid, recently shown to act as small-molecular-weight phospho-donors in the process of phosphorylation of several response regulators. These findings suggest that AlgR may react with intermediary metabolites such as carbamoyl phosphate and acetyl phosphate, and that these processes may play a role in the control of mucoidy in P. aeruginosa.
authors.affiliation: Department of Microbiology, University of Texas Health Science Center, San Antonio, 78284-7758.
authors.forename: V
authors.initials: V
authors.lastname: Deretic
authors.forename: J H
authors.initials: JH
authors.lastname: Leveau
authors.forename: C D
authors.initials: CD
authors.lastname: Mohr
authors.forename: N S
authors.initials: NS
authors.lastname: Hibler
mesh_terms: D000577:Amides / Q000494:pharmacology; D001426:Bacterial Proteins / Q000378:metabolism*; D002221:Carbamyl Phosphate / Q000494:pharmacology; D005798:Genes, Bacterial; D005809:Genes, Regulator; D000071677:Histidine Kinase; D008565:Membrane Proteins / Q000378:metabolism; D000072236:Methyl-Accepting Chemotaxis Proteins; D010755:Organophosphates / Q000494:pharmacology; D010756:Phosphoric Acids / Q000494:pharmacology; D010766:Phosphorylation; D011494:Protein Kinases / Q000378:metabolism*; D011550:Pseudomonas aeruginosa / Q000235:genetics / Q000378:metabolism*; D014157:Transcription Factors / Q000378:metabolism*
chemical_list: D000577:Amides; D001426:Bacterial Proteins; D008565:Membrane Proteins; D000072236:Methyl-Accepting Chemotaxis Proteins; D010755:Organophosphates; D010756:Phosphoric Acids; D014157:Transcription Factors; C011632:acetyl phosphate; D002221:Carbamyl Phosphate; C011067:phosphoramidic acid; D011494:Protein Kinases; D000071677:Histidine Kinase