期刊
MUTAGENESIS
卷 28, 期 1, 页码 81-88出版社
OXFORD UNIV PRESS
DOI: 10.1093/mutage/ges056
关键词
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资金
- Ministry of Education, Culture, Sports, Science and Technology [B-22710068]
- Japan Health Science Foundation [H24-food-general-011]
- Grants-in-Aid for Scientific Research [22710068, 22510075, 22241016] Funding Source: KAKEN
Many chronic inflammatory conditions are associated with an increased risk of cancer development. At the site of inflammation, cellular DNA is damaged by hypochlorous acid (HOCl), a potent oxidant generated by myeloperoxidase. 8-Chloro-2-deoxyguanosine (8-Cl-dG) is a major DNA adduct formed by HOCl and has been detected from the liver DNA and urine of rats administered lipopolysaccharide in an inflammation model. Thus, the 8-Cl-dG lesion may be associated with the carcinogenesis of inflamed tissues. In this study, we explored the miscoding properties of the 8-Cl-dG adduct generated by human DNA polymerases (pols). Site-specifically modified oligodeoxynucleotide containing a single 8-Cl-dG was prepared and used as a template in primer extension reactions catalysed by human pol , ? or ?. Primer extension reactions catalysed by pol and ? in the presence of all four dNTPs were slightly retarded at the 8-Cl-dG site, while pol ? readily bypassed the lesion. The fully extended products were analysed to quantify the miscoding frequency and specificity of 8-Cl-dG using two-phased polyacrylamide gel electrophoresis (PAGE). During the primer extension reaction in the presence of four dNTPs, pol ? promoted one-base deletion (6.4%), accompanied by the misincorporation of 2-deoxyguanosine monophosphate (5.5%), dAMP (3.7%), and dTMP (3.5%) opposite the lesion. Pol and ?, on the other hand, exclusively incorporated dCMP opposite the lesion. The steady-state kinetic studies supported the results obtained from the two-phased PAGE assay. These results indicate that 8-Cl-dG is a mutagenic lesion; the miscoding frequency and specificity varies depending on the DNA polymerase used. Thus, HOCl-induced 8-Cl-dG adduct may be involved in inflammation-driven carcinogenesis.
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