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Appl Environ Microbiol, February 1998, p. 411-418, Vol. 64, No. 2
0099-2240/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.

Expression and Regulation of the Arsenic Resistance Operon of Acidiphilium multivorum AIU 301 Plasmid pKW301 in Escherichia coli

Katsuhisa Suzuki,1 Norio Wakao,2 Tetsuya Kimura,1 Kazuo Sakka,1 and Kunio Ohmiya1,*

Laboratory of Applied Microbiology, School of Bioresources, Mie University, Tsu 514,1 and Laboratory of Applied Microbiology, Faculty of Agriculture, Iwate University, Morioka 020,2 Japan

Received 16 June 1997/Accepted 9 November 1997

The arsenic resistance (ars) operon from plasmid pKW301 of Acidiphilium multivorum AIU 301 was cloned and sequenced. This DNA sequence contains five genes in the following order: arsR, arsD, arsA, arsB, arsC. The predicted amino acid sequences of all of the gene products are homologous to the amino acid sequences of the ars gene products of Escherichia coli plasmid R773 and IncN plasmid R46. The ars operon cloned from A. multivorum conferred resistance to arsenate and arsenite on E. coli. Expression of the ars genes with the bacteriophage T7 RNA polymerase-promoter system allowed E. coli to overexpress ArsD, ArsA, and ArsC but not ArsR or ArsB. The apparent molecular weights of ArsD, ArsA, and ArsC were 13,000, 64,000, and 16,000, respectively. A primer extension analysis showed that the ars mRNA started at a position 19 nucleotides upstream from the arsR ATG in E. coli. Although the arsR gene of A. multivorum AIU 301 encodes a polypeptide of 84 amino acids that is smaller and less homologous than any of the other ArsR proteins, inactivation of the arsR gene resulted in constitutive expression of the ars genes, suggesting that ArsR of pKW301 controls the expression of this operon.

* Corresponding author. Mailing address: Laboratory of Applied Microbiology, School of Bioresources, Mie University, 1515 Kamihama-cho, Tsu 514, Japan. Phone and fax: 81-592-9622/9634. E-mail: ohmiya{at}bio.mie-u.ac.jp.




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