Metronidazole Activation Is Mutagenic and Causes DNA Fragmentation in Helicobacter pylori and in Escherichia coli Containing a Cloned H. pylori rdxA+ (Nitroreductase) Gene

doi:10.1128/JB.182.18.5091-5096.2000

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Journal of Bacteriology, September 2000, p. 5091-5096, Vol. 182, No. 18
0021-9193/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.

Metronidazole Activation Is Mutagenic and Causes DNA Fragmentation in Helicobacter pylori and in Escherichia coli Containing a Cloned H. pylori rdxA⁺ (Nitroreductase) Gene

Gary Sisson,¹ Jin-Yong Jeong,² Avery Goodwin,¹ Louis Bryden,¹ Norma Rossler,¹ Sabrina Lim-Morrison,¹ Ausra Raudonikiene,¹ Douglas E. Berg,² and Paul S. Hoffman¹^,³^,^*

Department of Microbiology and Immunology¹ and Division of Infectious Diseases, Department of Medicine,³ Dalhousie University, Halifax, Nova Scotia, Canada, and Department of Molecular Microbiology and Department of Genetics, Washington University School of Medicine, St. Louis, Missouri²

Received 23 March 2000/Accepted 28 June 2000

Much of the normal high sensitivity of wild-type Helicobacter pylori to metronidazole (Mtz) depends on rdxA (HP0954), a geneencoding a novel nitroreductase that catalyzes the conversionof Mtz from a harmless prodrug to a bactericidal agent. Here wereport that levels of Mtz that partially inhibit growth stimulateforward mutation to rifampin resistance in rdxA⁺ (Mtz^s) and also in rdxA (Mtz^r) H. pylori strains, and that expression of rdxA in Escherichiacoli results in equivalent Mtz-induced mutation. A reversion testusing defined lac tester strains of E. coli carrying rdxA⁺ indicated that CG-to-GC transversions and AT-to-GC transitionsare induced more frequently than other base substitutions. Alkalinegel electrophoretic tests showed that Mtz concentrations nearor higher than the MIC for growth also caused DNA breakage inH. pylori and in E. coli carrying rdxA⁺, suggesting that this damage may account for most of the bactericidalaction of Mtz. Coculture of Mtz^s H. pylori with E. coli (highly resistant to Mtz) in the presenceof Mtz did not stimulate forward mutation in E. coli, indicatingthat the mutagenic and bactericidal products of Mtz metabolismdo not diffuse significantly to neighboring (bystander) cells.Our results suggest that the widespread use of Mtz against otherpathogens in people chronically infected with H. pylori may stimulatemutation and recombination in H. pylori, thereby speeding host-specificadaptation, the evolution of virulence, and the emergence of resistanceagainst Mtz and other clinically usefulantimicrobials.

^* Corresponding author. Present address: Anti Infectives Research Division, SmithKline Beecham Pharmaceuticals, 1250 CollegevilleRoad, Collegeville, PA 19426. Phone: (610) 917-6010. Fax: (610)917-7901. E-mail: Paul_2_Hoffman{at}sbphrd.com or phoffman{at}tupdean2.med.dal.ca.

Journal of Bacteriology, September 2000, p. 5091-5096, Vol. 182, No. 18
0021-9193/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.

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