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Nucleic Acids Research 2005 33(7):2181-2191; doi:10.1093/nar/gki513
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Published online 14 April 2005

© The Author 2005. Published by Oxford University Press. All rights reserved
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Article

Repair activity of base and nucleotide excision repair enzymes for guanine lesions induced by nitrosative stress

Toshiaki Nakano, Atsushi Katafuchi, Ryoko Shimizu, Hiroaki Terato, Toshinori Suzuki1, Hiroshi Tauchi2, Keisuke Makino3, Milan Skorvaga4, Bennett Van Houten4 and Hiroshi Ide*

Department of Mathematical and Life Sciences, Graduate School of Science, Hiroshima University Higashi-Hiroshima 739-8526, Japan 1Department of Biological Pharmacy, School of Pharmacy, Shujitsu University 1-6-1 Nishigawara, Okayama 703-8516, Japan 2Department of Environmental Sciences, Faculty of Science, Ibaraki University Mito, Ibaraki 310-8512, Japan 3Institute of Advanced Energy, Kyoto University Gokasho, Uji 611-0011, Japan 4Laboratory of Molecular Genetics, National Institute of Environmental Health Sciences, National Institute of Health Research Triangle Park, NC 27709, USA

*To whom correspondence should be addressed. Tel: +81 82 424 7457; Fax: +81 82 424 7457; Email: ideh{at}hiroshima-u.ac.jp

Received January 26, 2005. Revised March 29, 2005. Accepted March 29, 2005.

Nitric oxide (NO) induces deamination of guanine, yielding xanthine and oxanine (Oxa). Furthermore, Oxa reacts with polyamines and DNA binding proteins to form cross-link adducts. Thus, it is of interest how these lesions are processed by DNA repair enzymes in view of the genotoxic mechanism of NO. In the present study, we have examined the repair capacity for Oxa and Oxa–spermine cross-link adducts (Oxa–Sp) of enzymes involved in base excision repair (BER) and nucleotide excision repair (NER) to delineate the repair mechanism of nitrosative damage to guanine. Oligonucleotide substrates containing Oxa and Oxa–Sp were incubated with purified BER and NER enzymes or cell-free extracts (CFEs), and the damage-excising or DNA-incising activity was compared with that for control (physiological) substrates. The Oxa-excising activities of Escherichia coli and human DNA glycosylases and HeLa CFEs were 0.2–9% relative to control substrates, implying poor processing of Oxa by BER. In contrast, DNA containing Oxa–Sp was incised efficiently by UvrABC nuclease and SOS-induced E.coli CFEs, suggesting a role of NER in ameliorating genotoxic effects associated with nitrosative stress. Analyses of the activity of CFEs from NER-proficient and NER-deficient human cells on Oxa–Sp DNA confirmed further the involvement of NER in the repair of nitrosative DNA damage.


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