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Journal of Bacteriology, September 2005, p. 5946-5954, Vol. 187, No. 17
0021-9193/05/$08.00+0     doi:10.1128/JB.187.17.5946-5954.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.

Transcriptional Termination Control of a Novel ABC Transporter Gene Involved in Antibiotic Resistance in Bacillus subtilis

Reiko Ohki,^* Kozue Tateno, Teruaki Takizawa, Toshiko Aiso, and Makiko Murata

Department of Molecular Biology, School of Health Sciences, Kyorin University, 476 Miyashita, Hachioji, Tokyo, 192-8508, Japan

Received 7 April 2005/ Accepted 7 June 2005

In members of one of the subfamilies of the bacterial ATP binding cassette (ABC) transporters, the two nucleotide binding domains are fused as a single peptide and the proteins have no membrane-spanning domain partners. Most of the ABC efflux transporters of this subfamily have been characterized in actinomycetes, producing macrolide, lincosamide, and streptogramin antibiotics. Among 40 ABC efflux transporters of Bacillus subtilis, five proteins belong to this subfamily. None of these proteins has been functionally characterized. We examined macrolide, lincosamide, and streptogramin antibiotic resistance in insertional disruptants of the genes that encode these proteins. It was found that only a disruptant of vmlR (formerly named expZ) showed hypersensitivity to virginiamycin M and lincomycin. Expression of the vmlR gene was induced by the addition of these antibiotics in growth medium. Primer extension analysis revealed that transcription of the vmlR gene initiates at an adenosine residue located 225 bp upstream of the initiation codon. From the analysis of the vmlR and lacZ fusion genes, a 52-bp deletion from +159 to +211 resulted in constitutive expression of the vmlR gene. In this region, a typical -independent transcriptional terminator was found. It was suggested that the majority of transcription ends at this termination signal in the absence of antibiotics, whereas under induced conditions, RNA polymerase reads through the terminator, and transcription continues to the downstream vmlR coding region, resulting in an increase in vmlR expression. No stabilization of vmlR mRNA occurred under the induced conditions.

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* Corresponding author. Mailing Address: Department of Molecular Biology, School of Health Sciences, Kyorin University, 476 Miyashita, Hachioji, Tokyo, 192-8508, Japan. Phone: 81 426 91 0011. Fax: 81 426 91 1094. E-mail: ohkir{at}kyorin-u.ac.jp.

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Journal of Bacteriology, September 2005, p. 5946-5954, Vol. 187, No. 17
0021-9193/05/$08.00+0     doi:10.1128/JB.187.17.5946-5954.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.

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