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Journal of Bacteriology, October 1998, p. 5484-5488, Vol. 180, No. 20
0021-9193/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.

Relationship between Spontaneous Aminoglycoside Resistance in Escherichia coli and a Decrease in Oligopeptide Binding Protein

Keiko Kashiwagi,1 Maria Heloisa Tsuhako,1 Kaori Sakata,1 Tomoko Saisho,1 Atsuko Igarashi,2 Sérgio Olavo Pinto da Costa,3 and Kazuei Igarashi1,*

Faculty of Pharmaceutical Sciences, Chiba University, Inage-ku, Chiba 263-8522,1 and Niigata University School of Dentistry, 2-5274 Gakkocho-dori, Niigata 951-8514,2 Japan, and Universidade de São Paulo, Instituto de Ciéncias Biomédicas, Departmento de Microbiologia, São Paulo 05508-900, Brazil3

Received 15 September 1997/Accepted 15 August 1998

Changes in the amount of oligopeptide binding protein (OppA) in spontaneous kanamycin-resistant mutants of Escherichia coli were investigated. Among 20 colonies obtained from 108 cells cultured in the presence of 20 µg of kanamycin/ml, 1 colony had no detectable OppA and 7 colonies were mutants with reduced amounts of OppA. Sensitivity of wild-type cells to kanamycin increased slightly by transformation of the oppA gene, but the sensitivity of the mutants increased greatly by the transformation. A mutant with no OppA was found to be a nonsense mutant of the oppA gene at amino acid position 166. In a mutant having a reduced level of OppA, the reduction was due to the decrease in OppA synthesis at the translational level. These mutants were also resistant to other aminoglycoside antibiotics, including streptomycin, neomycin, and isepamicin. Isepamicin uptake activities decreased greatly in these two kinds of mutants. The results support the proposition that aminoglycoside antibiotics are transported into cells by the oligopeptide transport system, and that transport is an important factor for spontaneous resistance to aminoglycoside antibiotics.

* Corresponding author. Mailing address: Faculty of Pharmaceutical Sciences, Chiba University, 1-33 Yayoi-cho, Inage-ku, Chiba 263-8522, Japan. Phone: 81-43-290-2897. Fax: 81-43-290-2900. E-mail: iga16077{at}p.chiba-u.ac.jp.

Journal of Bacteriology, October 1998, p. 5484-5488, Vol. 180, No. 20
0021-9193/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.


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