title: Porphyromonas gingivalis mutY is involved in the repair of oxidative stress-induced DNA mispairing.
abstract: The ability for DNA mismatch repair, after oxidative stress-induced DNA damage, is critical for the persistence of Porphyromonas gingivalis in the inflammatory environment of the periodontal pocket. Our previous report demonstrated that, in contrast to other organisms, the repair of oxidative stress-induced DNA damage involving 8-oxo-7,8-dihydroguanine (8-oxoG) may occur by a yet-to-be described mechanism in P. gingivalis. 8-oxoG does not block DNA replication; rather, it mispairs with adenine, which can be repaired by the MutY glycosylase. To determine the function of the P. gingivalis MutY homologue in DNA repair, it was insertionally inactivated using the ermF-ermAM antibiotic cassette and used to create a mutY-deficient mutant (FLL147) by allelic exchange mutagenesis. FLL147 had an increased rate of spontaneous mutation and was more sensitive to hydrogen peroxide compared with the wild-type W83 strain. DNA oligomers containing a site-specific 8-oxoG:A mispair was repaired similarly in both the P. gingivalis mutY-defective mutant and wild-type strains. The P. gingivalis mutY homologue was shown to complement the mutY mutation in Escherichia coli. In a gel mobility shift assay, the purified recombinant MutY is able to bind an oligo containing an 8-oxoG:A mispair. Taken together, MutY may play the expected role in oxidative stress resistance in P. gingivalis. However, there may exist other redundant mechanism(s) for the removal of 8-oxoG:A mismatch in this organism.
authors.affiliation: Division of Microbiology and Molecular Genetics, School of Medicine, Loma Linda University, Loma Linda, CA 92350, USA. agrobles@llu.edu
authors.forename: A G
authors.initials: AG
authors.lastname: Robles
authors.forename: K
authors.initials: K
authors.lastname: Reid
authors.forename: F
authors.initials: F
authors.lastname: Roy
authors.forename: H M
authors.initials: HM
authors.lastname: Fletcher
mesh_terms: D000225:Adenine; D001431:Bacteriological Techniques; D020137:Base Pair Mismatch / Q000235:genetics*; D001483:Base Sequence / Q000235:genetics; D017124:Conserved Sequence / Q000235:genetics; D004249:DNA Damage / Q000235:genetics; D045647:DNA Glycosylases / Q000235:genetics*; D053843:DNA Mismatch Repair / Q000235:genetics*; D005973:Glutamine / Q000235:genetics; D006147:Guanine / Q000031:analogs & derivatives / Q000032:analysis; D006801:Humans; D006861:Hydrogen Peroxide / Q000494:pharmacology; D016254:Mutagenesis, Insertional / Q000235:genetics; D016297:Mutagenesis, Site-Directed; D009154:Mutation / Q000235:genetics; D016877:Oxidants / Q000494:pharmacology; D018384:Oxidative Stress / Q000235:genetics*; D010957:Plasmids / Q000235:genetics; D016966:Porphyromonas gingivalis / Q000187:drug effects / Q000201:enzymology* / Q000235:genetics; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014633:Valine / Q000235:genetics
chemical_list: D016877:Oxidants; D005973:Glutamine; C024829:8-hydroxyguanine; D006147:Guanine; D006861:Hydrogen Peroxide; D045647:DNA Glycosylases; C061738:mutY adenine glycosylase; D014633:Valine; D000225:Adenine