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J. Biochem, 2004, Vol. 136, No. 3 359-362
© 2004 The Japanese Biochemical Society

MOLECULAR BIOLOGY

Increased A:T->C:G Mutations in the mutT Strain upon 8-Hydroxy-dGTP Treatment: Direct Evidence for MutT Involvement in the Prevention of Mutations by Oxidized dGTP

Hiroyuki Kamiya*, Chieko Ishiguro and Hideyoshi Harashima

Graduate School of Pharmaceutical Sciences, Hokkaido University, Kita-12, Nishi-6, Kita-ku, Sapporo 060-0812

The Escherichia coli MutT protein hydrolyzes 8-hydroxy-dGTP (8-OH-dGTP) in vitro, and mutT gene deficiencies cause increased spontaneous A:T->C:G mutations. However, no direct evidence exists for enhanced mutagenicity of 8-OH-dGTP in mutT cells. In this study, 8-OH-dGTP was introduced into wild type and mutT E. coli cells, and mutations of a chromosomal gene were monitored. 8-OH-dGTP induced mutations of the rpoB gene, the degree of the mutation induction in the mutT strain being ~6-fold higher than that in the wild type strain. On the other hand, 2-hydroxy-dATP, which is not a substrate of the MutT protein, increased the mutation to similar degrees in the two strains. These results constitute the first evidence that the MutT protein suppresses mutation by 8-OH-dGTP in vivo.

* To whom correspondence should be addressed. Tel: +81-11-706-3733, Fax: +81-11-706-4879, E-mail: hirokam{at}pharm.hokudai.ac.jp


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