Role of a MutY DNA glycosylase in combating oxidative DNA damage in Helicobacter pylori.
MutY is an adenine glycosylase that has the ability to efficiently remove adenines from adenine/7,8-dihydro-8-oxoguanine (8-oxo-G) or adenine/guanine mismatches, and plays an important role in oxidative DNA damage repair. The human gastric pathogen Helicobacter pylori has a homolog of the MutY enzyme. To investigate the physiological roles of MutY in H. pylori, we constructed and characterized a mutY mutant. H. pylori mutY mutants incubated at 5% O2 have a 325-fold higher spontaneous mutation rate than its parent. The mutation rate is further increased by exposing the mutant to atmospheric levels of oxygen, an effect that is not seen in an E. coli mutY mutant. Most of the mutations that occurred in H. pylori mutY mutants, as examined by rpoB sequence changes that confer rifampicin resistance, are GC to TA transversions. The H. pylori enzyme has the ability to complement an E. coli mutY mutant, restoring its mutation frequency to the wild-type level. Pure H. pylori MutY has the ability to remove adenines from A/8-oxo-G mismatches, but strikingly no ability to cleave A/G mismatches. This is surprising because E. coli MutY can more rapidly turnover A/G than A/8-oxo-G. Thus, H. pylori MutY is an adenine glycosylase involved in the repair of oxidative DNA damage with a specificity for detecting 8-oxo-G. In addition, H. pylori mutY mutants are only 30% as efficient as wild-type in colonizing the stomach of mice, indicating that H. pylori MutY plays a significant role in oxidative DNA damage repair in vivo.
Eutsey
Rory
R
University of Georgia, Athens, Georgia 30602, United States.
Wang
Ge
G
Maier
Robert J
RJ
D000595:Amino Acid Sequence; D000818:Animals; D004249:DNA Damage; D045647:DNA Glycosylases / Q000502:physiology*; D004260:DNA Repair; D005816:Genetic Complementation Test; D006147:Guanine / Q000737:chemistry / Q000378:metabolism; D016481:Helicobacter Infections / Q000201:enzymology / Q000235:genetics / Q000382:microbiology; D016480:Helicobacter pylori / Q000502:physiology*; D051379:Mice; D008810:Mice, Inbred C57BL; D008969:Molecular Sequence Data; D009154:Mutation; D018384:Oxidative Stress; D017386:Sequence Homology, Amino Acid; D013270:Stomach / Q000201:enzymology / Q000382:microbiology
D006147:Guanine; D045647:DNA Glycosylases; C061738:mutY adenine glycosylase