title: Mutagenesis and repair induced by the DNA advanced glycation end product N2-1-(carboxyethyl)-2'-deoxyguanosine in human cells.
abstract: Glycation of biopolymers by glucose-derived α-oxo-aldehydes such as methylglyoxal (MG) is believed to play a major role in the complex pathologies associated with diabetes and metabolic disease. In contrast to the extensive literature detailing the formation and physiological consequences of protein glycation, there is little information about the corresponding phenomenon for DNA. To assess the potential contribution of DNA glycation to genetic instability, we prepared shuttle vectors containing defined levels of the DNA glycation adduct N(2)-(1-carboxyethyl)-2'-deoxyguanosine (CEdG) and transfected them into isogenic human fibroblasts that differed solely in the capacity to conduct nucleotide excision repair (NER). In the NER-compromised fibroblasts, the induced mutation frequencies increased up to 18-fold relative to background over a range of ∼10-1400 CEdG adducts/10(5) dG, whereas the same substrates transfected into NER-competent cells induced a response that was 5-fold over background at the highest adduct density. The positive linear correlation (R(2) = 0.998) of mutation frequency with increasing CEdG level in NER-defective cells suggested that NER was the primary if not exclusive mechanism for repair of this adduct in human fibroblasts. Consistent with predictions from biochemical studies using CEdG-substituted oligonucleotides, guanine transversions were the predominant mutation resulting from replication of MG-modified plasmids. At high CEdG levels, significant increases in the number of AT → GC transitions were observed exclusively in NER-competent cells (P < 0.0001). This suggested the involvement of an NER-dependent mutagenic process in response to critical levels of DNA damage, possibly mediated by error-prone Y-family polymerases.
authors.affiliation: Division of Molecular Medicine, Beckman Research Institute of the City of Hope, 1500 East Duarte Road, Duarte, California 91010, United States.
authors.forename: Daniel
authors.initials: D
authors.lastname: Tamae
authors.forename: Punnajit
authors.initials: P
authors.lastname: Lim
authors.forename: Gerald E
authors.initials: GE
authors.lastname: Wuenschell
authors.forename: John
authors.initials: J
authors.lastname: Termini
mesh_terms: D001483:Base Sequence; D018736:DNA Adducts / Q000235:genetics* / Q000378:metabolism*; D004260:DNA Repair; D003849:Deoxyguanosine / Q000031:analogs & derivatives* / Q000378:metabolism; D005347:Fibroblasts / Q000378:metabolism*; D016153:Genes, Suppressor; D017127:Glycation End Products, Advanced / Q000378:metabolism*; D006801:Humans; D016296:Mutagenesis; D009154:Mutation; D010957:Plasmids / Q000235:genetics; D012343:RNA, Transfer / Q000235:genetics
chemical_list: D018736:DNA Adducts; D017127:Glycation End Products, Advanced; C400382:N(2)-(1-carboxyethyl)deoxyguanosine; C096275:supF tRNA; D012343:RNA, Transfer; D003849:Deoxyguanosine