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Nucleic Acids Research, 1995, Vol. 23, No. 5 761-766
© 1995

MOLECULAR BIOLOGY

Misincorporation of dAMP opposite 2-hydroxyadenine, an oxidative form of adenine

Hiroyuki Kamiya, Toshihiro Ueda1, Akio Matsukage2 and Hiroshi Kasai*

Department of Environmental Oncology, Institute of Industrial Ecological Sciences, University of Occupational and Environmental Health 1-1 Iseigaoka, Yahatanishi-ku, Kitakyushu 807, Japan 1Discovery Research Laboratory III, Nippon Shinyaku Co. Ltd Nishiohji-Hachijo, Minami-ku, Kyoto 601, Japan 2Laboratory of Cell Biology, Aichi Cancer Center Research Institute Chikusa-ku, Nagoya 464, Japan

*To whom correspondence should be addressed

Received November 21, 1994. Revised January 16, 1995. Accepted January 16, 1995.

Nucleotide incorporation opposite an oxidative form of adenine, 2-hydroxyadenine (2-OH-Ade) was investigated. When a primed template with 2-OH-Ade was treated with an exonuclease-deficient Klenow fragment of Escherichia coliDNA polymerase I (KFexo-), recombinant rat DNA polymerase ß (pol ß) or calf thymus DNA polymerase{alpha} (pol {alpha}), incorporation of dTMP and dAMP was observed. In addition, KFexo- inserted dGMP as well. A steady-state kinetic study indicated that the insertion of dAMP and dTMP opposite the DNA lesion occurred with similarfrequency with KFexo- and pol ß. Insertion of dTMP opposite 2-OH-Ade was favored to that of dAMP by pol {alpha}. Chain extension from the A-2-OH-Ade pair is less favored than that from the T-2-OH-Ade pair by all three DNA polymerases. Analysis of full-length products of in vitro DNA synthesis showed that dTMP and dAMP were Incorporated by DNA polymerases and that exonuclease-proficient and -deficient Klenow fragments also Inserted dGMP opposite 2-OH-Ade. These results suggest that formation of 2-OH-Ade from A in DNA will induce A->T and A->C transversions in cells.


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