title: RutR is the uracil/thymine-sensing master regulator of a set of genes for synthesis and degradation of pyrimidines.
abstract: Using the genomic SELEX, a total of six Escherichia coli DNA fragments have been identified, which formed complexes with transcription factor RutR. The RutR regulon was found to include a large number of genes encoding components for not only degradation of pyrimidines but also transport of glutamate, synthesis of glutamine, synthesis of pyrimidine nucleotides and arginine, and degradation of purines. DNase I footprinting indicated that RutR recognizes a palindromic sequence of TTGACCAnnTGGTCAA. The RutR box in P1 promoter of carAB encoding carbamoyl phosphate synthetase, a key enzyme of pyrimidine synthesis, overlaps with the PepA (CarP) repressor binding site, implying competition between RutR and PepA. Adding either uracil or thymine abolished RutR binding in vitro to the carAB P1 promoter. Accordingly, in the rutR-deletion mutant or in the presence of uracil, the activation in vivo of carAB P1 promoter was markedly reduced. Northern blot analysis of the RutR target genes indicated that RutR represses the Gad system genes involved in glutamate-dependent acid resistance and allantoin degradation. Altogether we propose that RutR is the pyrimidine sensor and the master regulator for a large set of the genes involved in the synthesis and degradation of pyrimidines.
authors.affiliation: Department of Frontier Bioscience and Micro-Nano Technology Research Centre, Hosei University, Koganei, Tokyo 184-8584, Japan.
authors.forename: Tomohiro
authors.initials: T
authors.lastname: Shimada
authors.forename: Kiyo
authors.initials: K
authors.lastname: Hirao
authors.forename: Ayako
authors.initials: A
authors.lastname: Kori
authors.forename: Kaneyoshi
authors.initials: K
authors.lastname: Yamamoto
authors.forename: Akira
authors.initials: A
authors.lastname: Ishihama
mesh_terms: D000481:Allantoin / Q000378:metabolism; D052157:Aptamers, Nucleotide / Q000235:genetics / Q000378:metabolism; D001483:Base Sequence; D001665:Binding Sites / Q000235:genetics; D015152:Blotting, Northern; D018983:DNA Footprinting / Q000379:methods; D003850:Deoxyribonuclease I / Q000378:metabolism; D024202:Electrophoretic Mobility Shift Assay; D029968:Escherichia coli Proteins / Q000235:genetics / Q000378:metabolism*; D015964:Gene Expression Regulation, Bacterial; D018698:Glutamic Acid / Q000378:metabolism; D008954:Models, Biological; D009154:Mutation; D011401:Promoter Regions, Genetic / Q000235:genetics; D011485:Protein Binding; D011743:Pyrimidines / Q000378:metabolism*; D052156:SELEX Aptamer Technique; D013941:Thymine / Q000378:metabolism*; D014157:Transcription Factors / Q000235:genetics / Q000378:metabolism*; D014498:Uracil / Q000378:metabolism*
chemical_list: D052157:Aptamers, Nucleotide; D029968:Escherichia coli Proteins; D011743:Pyrimidines; C530619:RutR protein, E coli; D014157:Transcription Factors; D000481:Allantoin; D018698:Glutamic Acid; D014498:Uracil; D003850:Deoxyribonuclease I; C030986:pyrimidine; D013941:Thymine