Abstract
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-CGG o, 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 η (Ypolη). 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 Ypolη 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 Ypolη. 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 η showed higher dATP incorporation rates even with the 5′-CCG o-3′ sequence.
| Original language | English |
|---|---|
| Pages (from-to) | 509-513 |
| Number of pages | 5 |
| Journal | Mutagenesis |
| Volume | 23 |
| Issue number | 6 |
| DOIs | |
| Publication status | Published - Nov 2008 |
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ASJC Scopus subject areas
- Genetics
- Genetics(clinical)
- Toxicology
- Health, Toxicology and Mutagenesis
Cite this
Influence of neighbouring base sequences on the mutagenesis induced by 7,8-dihydro-8-oxoguanine in yeast. / Yung, Chin Wei; Okugawa, Yoji; Otsuka, Chie; Okamoto, Keinosuke; Arimoto, Sakae; Loakes, David; Negishi, Kazuo; Negishi, Tomoe.
In: Mutagenesis, Vol. 23, No. 6, 11.2008, p. 509-513.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - Influence of neighbouring base sequences on the mutagenesis induced by 7,8-dihydro-8-oxoguanine in yeast
AU - Yung, Chin Wei
AU - Okugawa, Yoji
AU - Otsuka, Chie
AU - Okamoto, Keinosuke
AU - Arimoto, Sakae
AU - Loakes, David
AU - Negishi, Kazuo
AU - Negishi, Tomoe
PY - 2008/11
Y1 - 2008/11
N2 - 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-CGG o, 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 η (Ypolη). 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 Ypolη 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 Ypolη. 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 η showed higher dATP incorporation rates even with the 5′-CCG o-3′ sequence.
AB - 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-CGG o, 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 η (Ypolη). 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 Ypolη 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 Ypolη. 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 η showed higher dATP incorporation rates even with the 5′-CCG o-3′ sequence.
UR - http://www.scopus.com/inward/record.url?scp=55349138102&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=55349138102&partnerID=8YFLogxK
U2 - 10.1093/mutage/gen044
DO - 10.1093/mutage/gen044
M3 - Article
C2 - 18765421
AN - SCOPUS:55349138102
VL - 23
SP - 509
EP - 513
JO - Mutagenesis
JF - Mutagenesis
SN - 0267-8357
IS - 6
ER -