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Journal of Bacteriology, January 2003, p. 51-59, Vol. 185, No. 1
0021-9193/03/$08.00+0     DOI: 10.1128/JB.185.1.51-59.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.

A Bacitracin-Resistant Bacillus subtilis Gene Encodes a Homologue of the Membrane-Spanning Subunit of the Bacillus licheniformis ABC Transporter

Reiko Ohki,1* Kozue Tateno,1 Youji Okada,2 Haruo Okajima,2 Kei Asai,3 Yoshito Sadaie,3 Makiko Murata,1 and Toshiko Aiso1

Department of Molecular Biology,1 Department of Analytical Chemistry, School of Health Sciences, Kyorin University, Hachioji, Tokyo 192-0005,2 Department of Molecular Biology, Faculty of Science, Saitama University, 255 Shimoohkubo, Urawa, Saitama 338-8570, Japan3

Received 1 August 2002/ Accepted 2 October 2002

Bacitracin is a peptide antibiotic nonribosomally produced by Bacillus licheniformis. The bcrABC genes which confer bacitracin resistance to the bacitracin producer encode ATP binding cassette (ABC) transporter proteins, which are hypothesized to pump out bacitracin from the cells. Bacillus subtilis 168, which has no bacitracin synthesizing operon, has several genes homologous to bcrABC. It was found that the disruption of ywoA, a gene homologous to bcrC, resulted in hypersensitivity to bacitracin. Resistance to other drugs such as surfactin, iturin A, vancomycin, tunicamycin, gramicidin D, valinomycin and several cationic dyes were not changed in the ywoA disruptant. Spontaneous bacitracin-resistant mutants (Bcr-1 and -2) isolated in the presence of bacitracin have a single base substitution from A to G in the ribosome binding region. Northern hybridization analysis and determination of the expression of ywoA-LacZ transcriptional fusion gene revealed that the transcription of the ywoA gene was dependent on extracytoplasmic function (ECF) {sigma} factors {sigma}M and {sigma}X. Preincubation of wild-type cells in the presence of a low concentration of bacitracin induced increased resistance to bacitracin about two- to threefold, although the mechanism of this induction has not yet been elucidated. It has been reported that a commercially available bacitracin is a mixture of several components and also contains impurity. Bacitracin A was purified by reverse phase high-performance liquid chromatography (HPLC). Similar results were obtained with bacitracin A as those with crude bacitracin, indicating that contaminating substances were not responsible for the results obtained in this study.

* Corresponding author. Mailing address: Department of Molecular Biology, School of Health Sciences, Kyorin University, 476 Miyashita, Hachioji, Tokyo 192-0005, Japan. Phone: 81 426 91 0011. Fax: 81 426 91 1094. E-mail: ohkir{at}kyorin-u.ac.jp.

Journal of Bacteriology, January 2003, p. 51-59, Vol. 185, No. 1
0021-9193/03/$08.00+0     DOI: 10.1128/JB.185.1.51-59.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.



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