The major lipid peroxidation product, trans-4-hydroxy-2-nonenal, preferentially forms DNA adducts at codon 249 of human p53 gene, a unique mutational hotspot in hepatocellular carcinoma.
Trans-4-hydroxy-2-nonenal (4-HNE), a major electrophilic by-product of lipid peroxidation, is able to interact with DNA to form exocyclic guanine adducts. 4-HNE is a mutagen and a significant amount of 4-HNE-guanine adduct has been detected in normal cells. Recently, it has been reported that exposure of the wild-type p53 human lymphoblastoid cell line to 4-HNE causes a high frequency of G to T transversion mutations at the third base of codon 249 (-AGG*-) in the p53 gene, a mutational hotspot in human cancers, particularly hepatocellular carcinoma. These findings raise a possibility that 4-HNE could be an important etiological agent for human cancers that have a mutation at codon 249 of the p53 gene. However, to date, the sequence specificity of 4-HNE-DNA binding remains unclear due to the lack of methodology. To address this question, we have developed a method, using UvrABC nuclease, a nucleotide excision repair enzyme complex isolated from Escherichia coli, to map the distribution of 4-HNE-DNA adducts in human p53 gene at the nucleotide sequence level. We found that 4-HNE-DNA adducts are preferentially formed at the third base of codon 249 in the p53 gene. The preferential binding of 4-HNE was also observed at codon 174, which has the same sequence and the same nearest neighbor sequences (-GAGG*C-) as codon 249. These results suggest that 4-HNE may be an important etiological agent for human cancers that have a mutation at codon 249 of the p53 gene.
Hu
Wenwei
W
Department of Environmental Medicine, Pathology and Medicine, New York University School of Medicine, Tuxedo, NY 10987, USA.
Feng
Zhaohui
Z
Eveleigh
Jamie
J
Iyer
Ganesh
G
Pan
Jishen
J
Amin
Shantu
S
Chung
Fung-Lung
FL
Tang
Moon-Shong
MS
D000447:Aldehydes / Q000494:pharmacology*; D006528:Carcinoma, Hepatocellular / Q000235:genetics*; D002471:Cell Transformation, Neoplastic / Q000235:genetics; D003062:Codon / Q000235:genetics*; D018899:CpG Islands; D018736:DNA Adducts; D004249:DNA Damage; D019175:DNA Methylation; D004269:DNA, Bacterial / Q000187:drug effects / Q000378:metabolism; D004273:DNA, Neoplasm / Q000235:genetics*; D004278:DNA, Superhelical / Q000187:drug effects / Q000378:metabolism; D004706:Endodeoxyribonucleases / Q000378:metabolism; D029968:Escherichia coli Proteins / Q000378:metabolism; D016158:Genes, p53; D006801:Humans; D015227:Lipid Peroxidation; D008113:Liver Neoplasms / Q000235:genetics*; D009369:Neoplasms / Q000235:genetics; D010957:Plasmids / Q000187:drug effects / Q000235:genetics
D000447:Aldehydes; D003062:Codon; D018736:DNA Adducts; D004269:DNA, Bacterial; D004273:DNA, Neoplasm; D004278:DNA, Superhelical; D029968:Escherichia coli Proteins; D004706:Endodeoxyribonucleases; C036385:endodeoxyribonuclease uvrABC; C027576:4-hydroxy-2-nonenal