title: Cooperative regulation of the common target genes between H₂O₂-sensing YedVW and Cu²⁺-sensing CusSR in Escherichia coli.
abstract: YedVW is one of the uncharacterized two-component systems (TCSs) of Escherichia coli. In order to identify the regulation targets of YedVW, we performed genomic SELEX (systematic evolution of ligands by exponential enrichment) screening using phosphorylated YedW and an E. coli DNA library, and identified YedW-binding sites within three intergenic spacers, yedW-hiuH, cyoA-ampG and cusR-cusC, along the E. coli genome. Using a reporter assay system, we found that transcription of hiuH, encoding 5-hydroxyisourate hydrolase, was induced at high concentrations of either Cu(2+) or H₂O₂. Cu(2+)-dependent expression of hiuH was observed in the yedWV knockout mutant, but was reduced markedly in the cusRS-null mutant. However, H₂O₂-induced hiuH expression was observed in the cusRS-null mutant, but not in the yedWV-null mutant. Gel mobility shift and DNase I footprinting analyses showed binding of both YedW and CusR to essentially the same sequence within the hiuH promoter region. Taken together, we concluded that YedVW and CusSR formed a unique cooperative TCS pair by recognizing and regulating the same targets, but under different environmental conditions - YedVW played a role in H₂O₂ response regulation, whilst CusSR played a role in Cu(2+) response regulation.
authors.affiliation: Research Center for Human and Environmental Sciences, Shinshu University, Ueda, Nagano 386-8567, Japan.
authors.forename: Hiroyuki
authors.initials: H
authors.lastname: Urano
authors.affiliation: Department of Frontier Bioscience, Hosei University, Koganei, Tokyo 184-8584, Japan.
authors.forename: Yoshimasa
authors.initials: Y
authors.lastname: Umezawa
authors.affiliation: Department of Frontier Bioscience, Hosei University, Koganei, Tokyo 184-8584, Japan Research Center for Micro-Nano Technology, Hosei University, Koganei, Tokyo 184-8584, Japan.
authors.forename: Kaneyoshi
authors.initials: K
authors.lastname: Yamamoto
authors.affiliation: Department of Frontier Bioscience, Hosei University, Koganei, Tokyo 184-8584, Japan Research Center for Micro-Nano Technology, Hosei University, Koganei, Tokyo 184-8584, Japan.
authors.forename: Akira
authors.initials: A
authors.lastname: Ishihama
authors.affiliation: Research Center for Human and Environmental Sciences, Shinshu University, Ueda, Nagano 386-8567, Japan hogasawara@shinshu-u.ac.jp.
authors.forename: Hiroshi
authors.initials: H
authors.lastname: Ogasawara
mesh_terms: D001483:Base Sequence; D001665:Binding Sites; D003300:Copper / Q000378:metabolism*; D004926:Escherichia coli / Q000502:physiology*; D029968:Escherichia coli Proteins / Q000378:metabolism*; D015870:Gene Expression; D015964:Gene Expression Regulation, Bacterial; D017930:Genes, Reporter; D006861:Hydrogen Peroxide / Q000378:metabolism*; D011401:Promoter Regions, Genetic; D011485:Protein Binding; D052156:SELEX Aptamer Technique
chemical_list: D029968:Escherichia coli Proteins; D003300:Copper; D006861:Hydrogen Peroxide