Translesion synthesis of 7,8-dihydro-8-oxo-2′-deoxyguanosine by DNA polymerase eta in vivo
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7,8-Dihydro-8-oxo-2'-deoxyguanosine (8-oxo-dG) is one of the most common DNA lesions induced by oxidative stress. This lesion can be bypassed by DNA polymerase eta (Pol eta) using in vitro translesion synthesis (TLS) reactions. However, the role that Pol eta plays in vivo contributing to 8-oxo-dG mutagenesis remains unclear. To clarify the role of Pol eta in 8-oxo-dG mutagenesis, we have used an siRNA knockdown approach in combination with a supF shuttle vector (pSP189) which replicates in mammalian cells. The pSP189 plasmid was treated with methylene blue plus light (MBL), which produces predominantly 8-oxo-dG in DNA, and was then replicated in GM637 cells in presence of siRNA that knocks down the expression of Pol eta, or in XP-V cells, which lack functional Pol eta. The mutant frequencies were increased in the Pol eta siRNA knockdown cells and in XP-V cells relative to control, meaning that Pol eta plays an important role in preventing 8-oxo-dG mutagenesis. In the same system, knockdown of OGG1 also led to an increase in mutagenesis. Neither the type of mutations nor their distribution along the supF gene were significantly different between control and target specific siRNA-transfected cells (or XP-V cells) and were predominantly G to T transversions. These results show that Pol eta has an important role in error-free 8-oxo-dG lesion bypass and avoidance of oxidative stress-induced mutagenesis in vivo.
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G:C → T:A mutations are a signature of mutagenesis induced by 8-oxo-dG, the most common oxidative lesion in cells.
“Because G:C → T:A mutations are a signature of mutagenesis induced by 8-oxo-dG, the most common oxidative lesion in cells (37), this observation alludes to CSB ' s additional role in oxidative DNAdamage repair (21 -23), loss of which could result in increased oxidative damage-induced mutagenesis.”
From Why Cockayne syndrome patients do not get cancer despite their DNA repair deficiency (2016) · cites this paper as [37]
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- Authors
- Lee, D.-H., & Pfeifer, G. P.
- Affiliations
- Beckman Research Institute (US)
- Corresponding author
- Pfeifer, Gerd P.
- Venue
- Mutation Research - Fundamental and Molecular Mechanisms of Mutagenesis
- Publisher
- Elsevier BV
- Published
- 2008-05
- Volume
- 641
- Issue
- 1-2
- Pages
- 19-26
- ISSN
- 0027-5107 (ISSN-L)
- 1386-1964
- PMC
- PMC2410043
- OpenAlex
- W2076924472
- Cited by
- 53
- References
- 0
- Citations by year
- 2026: 1 · 2025: 1 · 2024: 1 · 2022: 1 · 2021: 1 · 2020: 1 · 2019: 2 · 2018: 2 · 2017: 5 · 2016: 3 · 2015: 3 · 2014: 6 · 2013: 4 · 2012: 6
- FWCI
- 2.19
- Type
- journal article
- Language
- en
- Open access
- green
- europepmc.org
- Licence
- Locations
- Mutation research. Fundamental and molecular mechanisms of mutagenesis (publishedVersion)
- PubMed (publishedVersion)
- Europe PMC (PubMed Central) (acceptedVersion)
- PubMed Central (acceptedVersion)
- Funders
- National Institutes of Health (ES06070)
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › DNA Repair Mechanisms
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Cancer Research › Carcinogens and Genotoxicity Assessment
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › DNA and Nucleic Acid Chemistry
- Keywords
- DNA polymerase eta, translesion synthesis, oxidative DNA damage, mutagenesis, OGG1, G to T transversions, lesion bypass
- MeSH terms
- 8-Hydroxy-2'-Deoxyguanosine, Y-Family DNA Polymerases, Base Sequence, Cells, Cultured, Deoxyguanosine, DNA, DNA Damage, DNA-Directed DNA Polymerase, DNA Repair, DNA Replication, Escherichia coli, Fibroblasts, Genetic Vectors, Humans, Light, Methylene Blue, Molecular Sequence Data, Skin, Transfection, Xeroderma Pigmentosum, Blotting, Western, Mutagenesis, Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization, Nucleic Acid Synthesis Inhibitors, Escherichia coli Proteins, RNA, Small Interfering, DNA Glycosylases
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