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Antimicrobial Agents and Chemotherapy, November 1998, p. 2906-2913, Vol. 42, No. 11
0066-4804/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.

Mutations in Bartonella bacilliformis gyrB Confer Resistance to Coumermycin A1

James M. Battisti, Laura S. Smitherman, D. Scott Samuels, and Michael F. Minnick*

Division of Biological Sciences, The University of Montana, Missoula, Montana 59812-1002

Received 17 April 1998/Returned for modification 1 June 1998/Accepted 13 August 1998

This study describes the first isolation and characterization of spontaneous mutants conferring natural resistance to an antibiotic for any Bartonella species. The Bartonella bacilliformis gyrB gene, which encodes the B subunit of DNA gyrase, was cloned and sequenced. The gyrB open reading frame (ORF) is 2,079 bp and encodes a deduced amino acid sequence of 692 residues, corresponding to a predicted protein of ~77.5 kDa. Sequence alignment indicates that B. bacilliformis GyrB is most similar to the GyrB protein from Bacillus subtilis (40.1% amino acid sequence identity) and that it contains the longest N-terminal tail (52 residues) of any GyrB characterized to date. The cloned B. bacilliformis gyrB was expressed in an Escherichia coli S30 cell extract and was able to functionally complement a temperature-sensitive E. coli Cour gyrB mutant (strain N4177). We isolated and characterized spontaneous mutants of B. bacilliformis resistant to coumermycin A1, an antibiotic that targets GyrB. Sequence analysis of gyrB from 12 Cour mutants of B. bacilliformis identified single nucleotide transitions at three separate loci in the ORF. The predicted amino acid substitutions resulting from these transitions are Gly to Ser at position 124 (Gly124right-arrowSer), Arg184right-arrowGln, and Thr214right-arrowAla or Thr214right-arrowIle, which are analogous to mutated residues found in previously characterized resistant gyrB genes from Borrelia burgdorferi, E. coli, Staphylococcus aureus, and Haloferax sp. The Cour mutants are three to five times more resistant to coumermycin A1 than the wild-type parental strain.

* Corresponding author. Mailing address: Division of Biological Sciences, The University of Montana, Missoula, MT 59812-1002. Phone: (406) 243-5972. Fax: (406) 243-4184. E-mail: minnick{at}selway.umt.edu.

Antimicrobial Agents and Chemotherapy, November 1998, p. 2906-2913, Vol. 42, No. 11
0066-4804/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.


This article has been cited by other articles:

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