Miscoding properties of 8-chloro-2′-deoxyguanosine, a hypochlorous acid-induced DNA adduct, catalysed by human DNA polymerases

Akira Sassa, Nagisa Kamoshita, Tomonari Matsuda, Yuji Ishii, Isao Kuraoka, Takehiko Nohmi, Toshihiro Ohta, Masamitsu Honma, Manabu Yasui*


研究成果: Article査読

14 被引用数 (Scopus)


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 α, κ or η. Primer extension reactions catalysed by pol α and κ in the presence of all four dNTPs were slightly retarded at the 8-Cl-dG site, while pol η 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 κ 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 α and η, 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.

出版ステータスPublished - 2013 1月

ASJC Scopus subject areas

  • 遺伝学
  • 毒物学
  • 遺伝学(臨床)
  • 健康、毒物学および変異誘発


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