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Antimicrobial Agents and Chemotherapy, December 2003, p. 3733-3738, Vol. 47, No. 12
0066-4804/03/$08.00+0     DOI: 10.1128/AAC.47.12.3733-3738.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.

EfrAB, an ABC Multidrug Efflux Pump in Enterococcus faecalis

Eun-Woo Lee, M. Nazmul Huda, Teruo Kuroda, Tohru Mizushima, and Tomofusa Tsuchiya*

Department of Microbiology, Faculty of Pharmaceutical Sciences, Okayama University, Tsushima, Okayama, 700-8530, Japan

Received 24 April 2003/ Returned for modification 28 July 2003/ Accepted 18 September 2003

A DNA fragment responsible for resistance to antimicrobial agents was cloned from the chromosomal DNA of Enterococcus faecalis ATCC 29212 by using drug-hypersensitive mutant Escherichia coli KAM32 as a host cell. Cells of E. coli KAM32 harboring a recombinant plasmid (pAEF82) carrying the DNA fragment became resistant to many structurally unrelated antimicrobial agents, such as norfloxacin, ciprofloxacin, doxycycline, acriflavine, 4',6-diamidino-2-phenylindole, tetraphenylphosphonium chloride, daunorubicin, and doxorubicin. Since the sequence of the whole genome of E. faecalis is known, we sequenced several portions of the DNA insert in plasmid pAEF82 and identified two open reading frames within the insert. We designated the genes efrA and efrB. A search of the deduced amino acid sequences of EfrA and EfrB revealed that they are similar to each other and that they belong to the ATP-binding cassette (ABC) family of multidrug efflux transporters. Transformed E. coli KAM32 cells harboring efrAB showed energy-dependent efflux of acriflavine. The efflux activity was inhibited by reserpine, verapamil, and sodium-o-vanadate, known inhibitors of ABC efflux pumps.

* Corresponding author. Mailing address: Department of Microbiology, Faculty of Pharmaceutical Sciences, Okayama University, Tsushima, Okayama, 700-8530, Japan. Phone and Fax: 81-86-251-7957. E-mail: tsuchiya{at}pharm.okayama-u.ac.jp.

Antimicrobial Agents and Chemotherapy, December 2003, p. 3733-3738, Vol. 47, No. 12
0066-4804/03/$08.00+0     DOI: 10.1128/AAC.47.12.3733-3738.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.



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