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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 gene encoding a novel nitroreductase that catalyzes the
conversion of Mtz from a harmless prodrug to a bactericidal agent. Here
we report that levels of Mtz that partially inhibit growth stimulate forward mutation to rifampin resistance in
rdxA+ (Mtzs) and also in
rdxA (Mtzr) H. pylori strains, and
that expression of rdxA in Escherichia coli
results in equivalent Mtz-induced mutation. A reversion test using
defined lac tester strains of E. coli carrying
rdxA+ indicated that CG-to-GC transversions and
AT-to-GC transitions are induced more frequently than other base
substitutions. Alkaline gel electrophoretic tests showed that Mtz
concentrations near or higher than the MIC for growth also caused DNA
breakage in H. pylori and in E. coli carrying
rdxA+, suggesting that this damage may account
for most of the bactericidal action of Mtz. Coculture of
Mtzs H. pylori with E. coli (highly
resistant to Mtz) in the presence of Mtz did not stimulate forward
mutation in E. coli, indicating that the mutagenic and
bactericidal products of Mtz metabolism do not diffuse significantly to
neighboring (bystander) cells. Our results suggest that the widespread
use of Mtz against other pathogens in people chronically infected with
H. pylori may stimulate mutation and recombination in
H. pylori, thereby speeding host-specific adaptation, the
evolution of virulence, and the emergence of resistance against Mtz and
other clinically useful antimicrobials.
*
Corresponding author. Present address: Anti Infectives
Research Division, SmithKline Beecham Pharmaceuticals, 1250 Collegeville Road, 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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