DNA replication fidelity with 8-oxodeoxyguanosine triphosphate
Abstract
Oxidative metabolism is known to generate mutagenic compounds within cells, among which is 8-oxodeoxyguanosine. Here the mutagenic potential of the triphosphate form of this base analog (8-O-dGTP) is investigated during replication in vitro of the lacZ alpha-complementation sequence in M13mp2 DNA. Adding 8-O-dGTP at equimolar concentration with the normal dNTPs to polymerization reactions decreases the fidelity of DNA synthesis by exonuclease-deficient Klenow, T4, and Thermus thermophilus DNA polymerases. Sequence analysis of mutants suggests that 8-O-dGMP is misincorporated opposite template adenines, yielding A-->C transversions. The degree of polymerase selectivity against this error is enzyme-dependent, with rates varying by > 25-fold. To determine if the A.8-O-dGMP mispair is proofread, a direct comparison of the fidelity of proofreading-proficient and proofreading-deficient Klenow and T4 DNA polymerases was made. Although the exonuclease activity of Klenow polymerase did not substantially reduce overall misincorporation of 8-O-dGMP, misincorporation was lower for the proofreading-proficient T4 enzyme as compared to its proofreading-deficient derivative. These data suggest that the A.8-O-dGMP mispair can be proofread. The mutagenic potential of 8-O-dGTP with eukaryotic systems was also examined. Misincorporation of 8-O-dGTP opposite adenine was observed during SV40 origin-dependent replication of double-stranded DNA in HeLa cell extracts. When present during replication at a concentration equal to the four normal dNTPs, 8-O-dGTP was at least 13-fold more mutagenic for A.T-->C.G transversions than was a 100-fold excess of normal dGTP.(ABSTRACT TRUNCATED AT 250 WORDS)
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MtDNA oxidation renders mtDNA more susceptible to mutation because oxidized bases are misread during replication and lead to nucleotide substitutions.
“MtDNA oxidation, in turn, renders mtDNA more susceptible to mutation because oxidized bases are misread during replication and this leads to nucleotide substitutions [118–120].”
From Brain aging, Alzheimer's disease, and mitochondria (2011) · cites this paper as [120]
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- Authors
- Pavlov, Y. I., Minnick, D. T., Izuta, S., & Kunkel, T. A.
- Affiliations
- National Institutes of Health (US)
- Venue
- Biochemistry
- Publisher
- American Chemical Society (ACS)
- Published
- 1994-04-19
- Volume
- 33
- Issue
- 15
- Pages
- 4695-4701
- ISSN
- 0006-2960 (ISSN-L)
- 1520-4995
- OpenAlex
- W1965072222
- Cited by
- 132
- References
- 43
- Citations by year
- 2026: 2 · 2025: 2 · 2024: 2 · 2021: 2 · 2020: 1 · 2019: 2 · 2018: 1 · 2017: 1 · 2016: 5 · 2015: 1 · 2014: 1 · 2013: 1
- FWCI
- 3.39
- Type
- journal article
- Language
- en
- Open access
- closed
- Locations
- Biochemistry (publishedVersion)
- PubMed (publishedVersion)
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › DNA Repair Mechanisms
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › DNA and Nucleic Acid Chemistry
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Cancer Research › Carcinogens and Genotoxicity Assessment
- Keywords
- 8-hydroxy-2'-deoxyguanosine, DNA replication fidelity, oxidative DNA damage, mutagenesis, misincorporation, DNA polymerase proofreading, exonuclease activity, Klenow fragment, T4 DNA polymerase, HeLa cell extracts, base analogs
- MeSH terms
- Base Composition, Deoxyguanine Nucleotides, DNA, DNA Polymerase I, DNA-Directed DNA Polymerase, DNA Replication, Escherichia coli, Glycosylation, Transfection, Mutagenesis, Thermus thermophilus, Bacteriophage T4
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