Identification and characterization of the zosA gene involved in copper uptake in Bacillus subtilis 168

Takahiro Fukuhara, Kazuo Kobayashi, Yousuke Kanayama, Shu Ichi Enomoto, Taeko Kondo, Naoki Tsunekawa, Michiko Nemoto, Naotake Ogasawara, Kenji Inagaki, Takashi Tamura

Research output: Contribution to journalArticle

2 Citations (Scopus)

Abstract

DL-Penicillamine, a copper-specific metal chelator, remarkably suppressed the growth of Bacillus subtilis 168 when added to a synthetic medium under Cu2+ 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 Cu2+ but not of excessive Zn2+ 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.

Original languageEnglish
Pages (from-to)600-609
Number of pages10
JournalBioscience, Biotechnology and Biochemistry
Volume80
Issue number3
DOIs
Publication statusPublished - 2016

Keywords

  • Bacillus subtilis 168
  • Cu-uptake ATPase
  • DL-penicillamine
  • Molecular dynamics simulation
  • ZosA

ASJC Scopus subject areas

  • Biotechnology
  • Biochemistry
  • Molecular Biology
  • Applied Microbiology and Biotechnology
  • Analytical Chemistry
  • Organic Chemistry

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