Using UvrABC nuclease to detect 7,12-dimethylbenz[a]anthracene anti- diol epoxide-DNA binding specificity in the mouse H-ras gene

James X. Chen, Alexander S. Kisleyou, Ronald G. Harvey, Thomas J. Slaga, Rebecca J. Morris, Moon Shong Tang

Research output: Contribution to journalArticlepeer-review

7 Scopus citations

Abstract

DNA fragments modified with chemically synthesized 7, 12- dimethylbenz[a]anthracene anti-diol epoxide (anti-DMBADE) are sensitive to UvrABC nuclease incision. The incisions occur mainly 7 bases 5' and 4 bases 3' of an anti-DMBADE-modified adenine or guanine residue, and the kinetics of incision at different sequences in a DNA fragment are the same. These results indicate that UvrABC incision on anti-DMBADE-DNA adducts is independent of DNA sequences and is quantitative, the same as on syn-DMBADE-DNA adducts. This method was used to analyze the anti-DMBADE-DNA binding spectrum in the exon 2 region of the mouse H-ras gene, and it was found that anti-DMBADE binds to the two adenine residues at codon 61 of the H-ras gene with an average affinity. Previously, we have demonstrated that syn-DMBADE binds strongly to the adenines at codon 61 of H-ras; these results together suggest that the oncogenic mutation in H-ras may be induced by anti- and syn-DMBADE- DNA adducts.

Original languageEnglish (US)
Pages (from-to)1350-1354
Number of pages5
JournalChemical research in toxicology
Volume9
Issue number8
DOIs
StatePublished - 1996
Externally publishedYes

ASJC Scopus subject areas

  • Toxicology

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