title: Repair of oxidative DNA damage is delayed in the Ser326Cys polymorphic variant of the base excision repair protein OGG1.
abstract: Gene-environment interactions influence an individual's risk of disease development. A common human 8-oxoguanine DNA glycosylase 1 (OGG1) variant, Cys326-hOGG1, has been associated with increased cancer risk. Evidence suggests that this is due to reduced repair ability, particularly under oxidising conditions but the underlying mechanism is poorly understood. Oxidising conditions may arise due to internal cellular processes, such as inflammation or external chemical or radiation exposure. To investigate wild-type and variant OGG1 regulation and activity under oxidising conditions, we generated mOgg1 (-/-) null mouse embryonic fibroblasts cells stably expressing Ser326- and Cys326-hOGG1 and measured activity, gene expression, protein expression and localisation following treatment with the glutathione-depleting compound L-buthionine-S-sulfoximine (BSO). Assessment of OGG1 activity using a 7,8-dihydro-8-oxodeoxyguanine (8-oxo dG) containing molecular beacon demonstrated that the activity of both Ser326- and Cys326-hOGG1 was increased following oxidative treatment but with different kinetics. Peak activity of Ser326-hOGG1 occurred 12 h prior to that of Cys326-hOGG1. In both variants, the increased activity was not associated with any gene expression or protein increase or change in protein localisation. These findings suggest that up-regulation of OGG1 activity in response to BSO-induced oxidative stress is via post-transcriptional regulation and provide further evidence for impaired Cys326-hOGG1 repair ability under conditions of oxidative stress. This may have important implications for increased mutation frequency resulting from increased oxidative stress in individuals homozygous for the Cys326 hOGG1 allele.
authors.affiliation: School of Biosciences, University of Birmingham, Vincent Drive, Edgbaston, Birmingham, B15 2TT, UK.
authors.forename: Rachael M
authors.initials: RM
authors.lastname: Kershaw
authors.forename: Nikolas J
authors.initials: NJ
authors.lastname: Hodges
mesh_terms: D000818:Animals; D000963:Antimetabolites / Q000494:pharmacology; D015153:Blotting, Western; D019328:Buthionine Sulfoximine / Q000494:pharmacology; D002478:Cells, Cultured; D003545:Cysteine / Q000737:chemistry / Q000235:genetics; D004249:DNA Damage / Q000235:genetics*; D045647:DNA Glycosylases / Q000235:genetics / Q000378:metabolism* / Q000502:physiology*; D004260:DNA Repair / Q000235:genetics*; D003839:Deoxyadenosines / Q000378:metabolism; D004622:Embryo, Mammalian / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D005347:Fibroblasts / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D005434:Flow Cytometry; D059647:Gene-Environment Interaction; D005978:Glutathione / Q000378:metabolism; D006801:Humans; D051379:Mice; D018345:Mice, Knockout; D059645:Mutation Rate; D018384:Oxidative Stress / Q000235:genetics*; D011110:Polymorphism, Genetic / Q000235:genetics*; D012333:RNA, Messenger / Q000235:genetics; D017382:Reactive Oxygen Species / Q000378:metabolism; D060888:Real-Time Polymerase Chain Reaction; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012694:Serine / Q000737:chemistry / Q000235:genetics; D015854:Up-Regulation
chemical_list: D000963:Antimetabolites; D003839:Deoxyadenosines; D012333:RNA, Messenger; D017382:Reactive Oxygen Species; D012694:Serine; D019328:Buthionine Sulfoximine; C078251:2'-deoxy-7,8-dihydro-8-oxoadenosine; D045647:DNA Glycosylases; C485990:Ogg1 protein, mouse; C479058:oxoguanine glycosylase 1, human; D005978:Glutathione; D003545:Cysteine