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Nucleic Acids Research, 2002, Vol. 30, No. 22 4975-4984
© 2002 Oxford University Press

Novel repair activities of AlkA (3-methyladenine DNA glycosylase II) and endonuclease VIII for xanthine and oxanine, guanine lesions induced by nitric oxide and nitrous acid

Hiroaki Terato, Aya Masaoka, Kenjiro Asagoshi, Akiko Honsho, Yoshihiko Ohyama, Toshinori Suzuki1, Masaki Yamada1, Keisuke Makino1, Kazuo Yamamoto2 and Hiroshi Ide*

Department of Mathematical and Life Sciences, Graduate School of Science, Hiroshima University, Higashi-Hiroshima 739-8526, Japan, 1 Institute of Advanced Energy, Kyoto University, Gokasho, Uji 611-0011, Japan and 2 Biological Institute, Graduate School of Science, Tohoku University, Sendai 980-8578, Japan

*To whom correspondence should be addressed. Tel/Fax: +81 824 24 7457; Email: ideh{at}hiroshima-u.ac.jp
Present address:
Toshinori Suzuki, Unit of Endogenous Cancer Risk Factors, International Agency for Research on Cancer, Lyon, France

Nitrosation of guanine in DNA by nitrogen oxides such as nitric oxide (NO) and nitrous acid leads to formation of xanthine (Xan) and oxanine (Oxa), potentially cytotoxic and mutagenic lesions. In the present study, we have examined the repair capacity of DNA N-glycosylases from Escherichia coli for Xan and Oxa. The nicking assay with the defined substrates containing Xan and Oxa revealed that AlkA [in combination with endonuclease (Endo) IV] and Endo VIII recognized Xan in the tested enzymes. The activity (Vmax/Km) of AlkA for Xan was 5-fold lower than that for 7-methylguanine, and that of Endo VIII was 50-fold lower than that for thymine glycol. The activity of AlkA and Endo VIII for Xan was further substantiated by the release of [3H]Xan from the substrate. The treatment of E.coli with N-methyl-N'-nitro-N-nitrosoguanidine increased the Xan-excising activity in the cell extract from alkA+ but not alkA– strains. The alkA and nei (the Endo VIII gene) double mutant, but not the single mutants, exhibited increased sensitivity to nitrous acid relative to the wild type strain. AlkA and Endo VIII also exhibited excision activity for Oxa, but the activity was much lower than that for Xan.


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