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Mutagenesis Advance Access originally published online on September 2, 2008
Mutagenesis 2008 23(6):509-513; doi:10.1093/mutage/gen044
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© The Author 2008. Published by Oxford University Press on behalf of the UK Environmental Mutagen Society. All rights reserved. For permissions, please e-mail: journals.permissions@oxfordjournals.org.

Influence of neighbouring base sequences on the mutagenesis induced by 7,8-dihydro-8-oxoguanine in yeast

Chin-Wei Yung, Yoji Okugawa, Chie Otsuka1, Keinosuke Okamoto, Sakae Arimoto, David Loakes2, Kazuo Negishi3 and Tomoe Negishi*

Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama University, Tsushima, Okayama 700-8530, Japan 1Department of Biochemistry, Shujitu University School of Pharmacy, 1-6-1 Nishigawara, Okayama 703-8516, Japan 2Medical Research Council, Laboratory of Molecular Biology, Hills Road, Cambridge CB2 2QH, UK 3Department of Immunobiology, Nihon Pharmaceutical University, 10281 Komuro, Ina, Saitama 362-0806, Japan

We have analysed the influence of neighbouring base sequences on the mutagenesis induced by 7,8-dihydro-8-oxoguanine (8-oxoG or Go), a typical oxidative lesion of DNA, using the yeast oligonucleotide transformation technique. Two oligonucleotides, oligo-CCGo and oligo-CGGo, each possessing a single 8-oxoG residue and represented by the sequences 5'-CCGo-3' and 5'-CGGo-3', respectively, were introduced into a chromosome of Saccharomyces cerevisiae and their mutagenic potentials were compared. In a wild-type strain, 8-oxoG showed very weak mutagenic potential in both cases. However, the lesion in 5'-CCGo-3' can cause efficient G-to-T transversion in a strain lacking the rad30 gene which encodes yeast DNA polymerase {eta} (Ypol{eta}). To explore the properties associated with this translesion synthesis (TLS), the same two oligonucleotides possessing an 8-oxoG were used as templates for a standing-start primer extension assay, and the nucleotide incorporation opposite 8-oxoG was investigated. We found that dATP incorporation opposite 8-oxoG with Ypol{eta} was low for both sequences. In particular, very low dATP incorporation was observed for the 5'-CCGo-3' sequence. These results account for the efficient inhibition of mutagenesis by Ypol{eta}. TLS plays an important role in one DNA sequence in terms of avoiding mutagenesis induced by 8-oxoG in yeast. In contrast, human yeast DNA polymerase {eta} showed higher dATP incorporation rates even with the 5'-CCGo-3' sequence.

* To whom correspondence should be addressed. Tel: +81 86 251 7946; Fax: +81 86 251 7926; Email: isaka{at}pheasant.pharm.okayama-u.ac.jp

Received on May 5, 2008; revised on July 16, 2008; accepted on July 28, 2008.


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S. V. Mudrak, C. Welz-Voegele, and S. Jinks-Robertson
The Polymerase {eta} Translesion Synthesis DNA Polymerase Acts Independently of the Mismatch Repair System To Limit Mutagenesis Caused by 7,8-Dihydro-8-Oxoguanine in Yeast
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[Abstract] [Full Text] [PDF]


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