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Journal of Bacteriology, September 2002, p. 5027-5035, Vol. 184, No. 18
0021-9193/02/$04.00+0     DOI: 10.1128/JB.184.18.5027-5035.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.

A Novel Histidine-Rich CPx-ATPase from the Filamentous Cyanobacterium Oscillatoria brevis Related to Multiple-Heavy-Metal Cotolerance

Liu Tong, Susumu Nakashima, Mineo Shibasaka, Maki Katsuhara, and Kunihiro Kasamo*

Research Institute for Bioresources, Okayama University, Kurashiki, Okayama 710-0046, Japan

Received 22 April 2002/ Accepted 21 June 2002

A novel gene related to heavy-metal transport was cloned and identified from the filamentous cyanobacterium Oscillatoria brevis. Sequence analysis of the gene (the Bxa1 gene) showed that its product possessed high homology with heavy-metal transport CPx-ATPases. The CPC motif, which is proposed to form putative cation transduction channel, was found in the sixth transmembrane helix. However, instead of the CXXC motif that is present in the N termini of most metal transport CPx-ATPases, Bxa1 contains a unique Cys-Cys (CC) sequence element and histidine-rich motifs as a putative metal binding site. Northern blotting and real-time quantitative reverse transcription-PCR showed that expression of Bxa1 mRNA was induced in vivo by both monovalent (Cu+ and Ag+) and divalent (Zn2+ and Cd2+) heavy-metal ions at similar levels. Experiments on heavy-metal tolerance in Escherichia coli with recombinant Bxa1 demonstrated that Bxa1 conferred resistance to both monovalent and divalent heavy metals. This is the first report of a CPx-ATPase responsive to both monovalent and divalent heavy metals.

* Corresponding author. Mailing address: Research Institute for Bioresources, Okayama University, Kurashiki, Okayama 710-0046, Japan. Phone: 81-86-434-1221. Fax: 81-86-434-1221. E-mail: kasamo{at}rib.okayama-u.ac.jp.

Journal of Bacteriology, September 2002, p. 5027-5035, Vol. 184, No. 18
0021-9193/02/$04.00+0     DOI: 10.1128/JB.184.18.5027-5035.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.



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