title: Identification and characterization of the zosA gene involved in copper uptake in Bacillus subtilis 168.
abstract: DL-Penicillamine, a copper-specific metal chelator, remarkably suppressed the growth of Bacillus subtilis 168 when added to a synthetic medium under Cu(2+) limitation. DNA microarray and screening of 2,602 knockout mutants showed that the zosA gene was de-repressed in the presence of 0.1% dl-penicillamine, and that the zosA mutant was sensitive to dl-penicillamine medium. The zosA mutant delayed the growth under Cu-limitation even without the chelator, and the sensitivity to dl-penicillamine was reversed by induction using 0.3 mM IPTG and the Pspac promoter inserted directly upstream of the zosA gene. Furthermore, the zosA mutant showed elevated tolerance of excessive Cu(2+) but not of excessive Zn(2+) added to LB and synthetic media. Homology modeling of the ZosA protein suggested that the protein can fold itself into essential domains for constituting a metal transporting ATPase. Our study suggests that zosA is a candidate gene involved in copper uptake.
authors.affiliation: a Graduate School of Life and Environmental Science , Okayama University , Okayama , Japan.
authors.forename: Takahiro
authors.initials: T
authors.lastname: Fukuhara
authors.affiliation: b Graduate School of Information Science , Nara Institute of Science & Technology , Ikoma , Japan.
authors.forename: Kazuo
authors.initials: K
authors.lastname: Kobayashi
authors.affiliation: c Laboratory of Multiple Molecular Imaging Research , Center for Molecular Imaging Science, RIKEN Kobe Institute , Hyogo , Japan.
authors.forename: Yousuke
authors.initials: Y
authors.lastname: Kanayama
authors.affiliation: c Laboratory of Multiple Molecular Imaging Research , Center for Molecular Imaging Science, RIKEN Kobe Institute , Hyogo , Japan.
authors.forename: Shu-ichi
authors.initials: S
authors.lastname: Enomoto
authors.affiliation: a Graduate School of Life and Environmental Science , Okayama University , Okayama , Japan.
authors.forename: Taeko
authors.initials: T
authors.lastname: Kondo
authors.affiliation: d Institute of Molecular and Cellular Biosciences , The University of Tokyo , Tokyo , Japan.
authors.forename: Naoki
authors.initials: N
authors.lastname: Tsunekawa
authors.affiliation: a Graduate School of Life and Environmental Science , Okayama University , Okayama , Japan.
authors.forename: Michiko
authors.initials: M
authors.lastname: Nemoto
authors.affiliation: b Graduate School of Information Science , Nara Institute of Science & Technology , Ikoma , Japan.
authors.forename: Naotake
authors.initials: N
authors.lastname: Ogasawara
authors.affiliation: a Graduate School of Life and Environmental Science , Okayama University , Okayama , Japan.
authors.forename: Kenji
authors.initials: K
authors.lastname: Inagaki
authors.affiliation: a Graduate School of Life and Environmental Science , Okayama University , Okayama , Japan.|e PRESTO, Japan Science and Technology Agency , Tokyo , Japan.
authors.forename: Takashi
authors.initials: T
authors.lastname: Tamura
mesh_terms: D001412:Bacillus subtilis / Q000235:genetics* / Q000378:metabolism; D003300:Copper / Q000378:metabolism*; D005798:Genes, Bacterial; D009154:Mutation; D020411:Oligonucleotide Array Sequence Analysis
chemical_list: D003300:Copper
keywords: Bacillus subtilis 168; Cu-uptake ATPase; dl-penicillamine; molecular dynamics simulation; zosA