title: Effect of CpG methylation at different sequence context on acrolein- and BPDE-DNA binding and mutagenesis.
abstract: Acrolein (Acr), an α,β-unsaturated aldehyde, is abundant in tobacco smoke and cooking and exhaust fumes. Acr induces mutagenic α- and γ- hydroxy-1,N(2)-cyclic propano-deoxyguanosine adducts in normal human bronchial epithelial cells. Our earlier work has found that Acr-induced DNA damage preferentially occurs at lung cancer p53 mutational hotspots that contain CpG sites and that methylation at CpG sites enhances Acr-DNA binding at these sites. Based on these results, we hypothesized that this enhancement of Acr-DNA binding leads to p53 mutational hotspots in lung cancer. In this study, using a shuttle vector supF system, we tested this hypothesis by determining the effect of CpG methylation on Acr-DNA binding and the mutations in human lung fibroblasts. We found that CpG methylation enhances Acr-induced mutations significantly. Although CpG methylation enhances Acr-DNA binging at all CpG sites, it enhances mutations at selective--TCGA--sites. Similarly, we found that CpG methylation enhances benzo(a)pyrene diol epoxide binding at all -CpG- sites. However, the methylated CpG sequences in which benzo(a)pyrene diol epoxide-induced mutations are enhanced are different from the CpG sequences in which Acr-induced mutations are enhanced. CpG methylation greatly increases Acr-induced G to T and G to A mutation frequency to levels similar to these types of mutations found in the CpG sites in the p53 gene in tobacco smoke-related lung cancer. These results indicate that both CpG sequence context and the chemical nature of the carcinogens are crucial factors for determining the effect of CpG methylation on mutagenesis.
authors.affiliation: Department of Environmental Medicine, Pathology and Medicine, New York University School of Medicine, Tuxedo Park, New York 10987, USA.
authors.forename: Hsiang-Tsui
authors.initials: HT
authors.lastname: Wang
authors.forename: Mao-wen
authors.initials: MW
authors.lastname: Weng
authors.forename: Wen-chi
authors.initials: WC
authors.lastname: Chen
authors.forename: Michael
authors.initials: M
authors.lastname: Yobin
authors.forename: Jishen
authors.initials: J
authors.lastname: Pan
authors.forename: Fung-Lung
authors.initials: FL
authors.lastname: Chung
authors.forename: Xue-Ru
authors.initials: XR
authors.lastname: Wu
authors.forename: William
authors.initials: W
authors.lastname: Rom
authors.forename: Moon-shong
authors.initials: MS
authors.lastname: Tang
mesh_terms: D015123:7,8-Dihydro-7,8-dihydroxybenzo(a)pyrene 9,10-oxide / Q000378:metabolism*; D000171:Acrolein / Q000378:metabolism / Q000633:toxicity*; D001483:Base Sequence; D002478:Cells, Cultured; D018899:CpG Islands; D004247:DNA / Q000187:drug effects / Q000235:genetics; D018736:DNA Adducts / Q000378:metabolism*; D019175:DNA Methylation; D017931:DNA Primers; D006801:Humans; D008969:Molecular Sequence Data; D009153:Mutagens / Q000378:metabolism / Q000633:toxicity*; D016133:Polymerase Chain Reaction
chemical_list: D018736:DNA Adducts; D017931:DNA Primers; D009153:Mutagens; C043931:benzo(a)pyrene-7,8-dihydrodiol-9,10-epoxide-DNA; D015123:7,8-Dihydro-7,8-dihydroxybenzo(a)pyrene 9,10-oxide; D000171:Acrolein; D004247:DNA