title: Characterization of fhlA mutations resulting in ligand-independent transcriptional activation and ATP hydrolysis.
abstract: The FhlA protein belongs to the NtrC family of transcriptional regulators. It induces transcription from the -12/-24 promoters of the genes of the formate regulon by sigma54 RNA polymerase. FhlA is activated by binding of the ligand formate and does not require phosphorylation. A mutational analysis of the fhLA gene portion coding for the A and C domains was conducted with the aim of gaining information on the interaction between formate binding and ATP hydrolysis plus transcription activation. Four mutations were identified, all located in the A domain; one of them rendered transcription completely independent from the presence of formate, and the others conferred a semiconstitutive phenotype. The FhlA protein of one of the semiconstitutive variants was purified. Catalytic efficiency of ATP hydrolysis of the mutant FhlA was increased in the absence of formate in the same manner as formate influences the activity of wild-type FhlA. Moreover, in vitro transcription occurred at much lower threshold concentrations of the mutant protein and of nucleoside triphosphates than with the wild-type FhlA.
authors.affiliation: Lehrstuhl für Mikrobiologie der Universität München, Munich, Germany.
authors.forename: I
authors.initials: I
authors.lastname: Korsa
authors.forename: A
authors.initials: A
authors.lastname: Böck
mesh_terms: D000255:Adenosine Triphosphate / Q000378:metabolism*; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D029968:Escherichia coli Proteins; D005561:Formates / Q000494:pharmacology*; D015964:Gene Expression Regulation, Bacterial / Q000187:drug effects*; D005798:Genes, Bacterial / Q000235:genetics; D006868:Hydrolysis; D007700:Kinetics; D008024:Ligands; D009154:Mutation; D015534:Trans-Activators / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism; D015533:Transcriptional Activation / Q000187:drug effects
chemical_list: D029968:Escherichia coli Proteins; D005561:Formates; D008024:Ligands; D015534:Trans-Activators; C030544:formic acid; C067189:fhlA protein, E coli; D000255:Adenosine Triphosphate