title: Involvement of the global Crp regulator in cyclic AMP-dependent utilization of aromatic amino acids by Pseudomonas putida.
abstract: The phhAB operon encodes a phenylalanine hydroxylase involved in the conversion of L-phenylalanine into L-tyrosine in Pseudomonas putida. The phhAB promoter is transcribed by RNA polymerase sigma-70 and is unusual in that the specific regulator PhhR acts as an enhancer protein that binds to two distant upstream sites (-75 to -92 and -132 to -149). There is an integration host factor (IHF) binding site that overlaps the proximal PhhR box, and, consequently, IHF acts as an inhibitor of transcription. Use of L-phenylalanine is compromised in a crp-deficient background due to reduced expression from the phhAB promoter. Electrophoretic mobility shift assays and DNase I footprinting assays reveal that Crp binds at a site centered at -109 only in the presence of cyclic AMP (cAMP). We show, using circular permutation analysis, that the simultaneous binding of Crp/cAMP and PhhR bends DNA to bring positive regulators and RNA polymerase into close proximity. This nucleoprotein complex promotes transcription from phhA only in response to L-phenylalanine.
authors.affiliation: Consejo Superior de Investigaciones Científicas, Estación Experimental del Zaidín, Department of Environmental Protection, Granada, Spain.
authors.forename: M Carmen
authors.initials: MC
authors.lastname: Herrera
authors.forename: Abdelali
authors.initials: A
authors.lastname: Daddaoua
authors.forename: Ana
authors.initials: A
authors.lastname: Fernández-Escamilla
authors.forename: Juan-Luis
authors.initials: JL
authors.lastname: Ramos
mesh_terms: D024322:Amino Acids, Aromatic / Q000378:metabolism*; D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism*; D003216:Conditioning, Operant; D000242:Cyclic AMP / Q000235:genetics / Q000378:metabolism*; D002373:Cyclic AMP Receptor Protein / Q000235:genetics / Q000378:metabolism*; D015964:Gene Expression Regulation, Bacterial / Q000502:physiology; D008958:Models, Molecular; D010649:Phenylalanine / Q000378:metabolism; D016958:Pseudomonas putida / Q000235:genetics* / Q000378:metabolism*; D014443:Tyrosine / Q000378:metabolism
chemical_list: D024322:Amino Acids, Aromatic; D001426:Bacterial Proteins; D002373:Cyclic AMP Receptor Protein; D014443:Tyrosine; D010649:Phenylalanine; D000242:Cyclic AMP
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