Cited by 133DNA Repair MechanismsDNA and Nucleic Acid ChemistryCarcinogens and Genotoxicity Assessment

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DNA Replication Fidelity with 8-Oxodeoxyguanosine Triphosphate

Pavlov, Youri I., Minnick, Dana T., Izuta, Shunji, Kunkel, Thomas A.·1994·Biochemistry

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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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Authors: Pavlov, Y. I., Minnick, D. T., Izuta, S., & Kunkel, T. A.
Affiliations: National Institutes of Health (US)
ORCID: Pavlov, Youri I.
Kunkel, Thomas A.
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
DOI: 10.1021/bi00181a029
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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DNA Replication Fidelity with 8-Oxodeoxyguanosine Triphosphate

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