Oxidative nucleotide damage: consequences and prevention

Sekiguchi, Mutsuo; Tsuzuki, Teruhisa

doi:10.1038/sj.onc.1206023

Cited by 203 publications

(172 citation statements)

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“…Currently, the prevailing paradigm is that 8-oxodGTP is predominantly formed directly from dGTP in cells and that there may be some contribution from 8-oxodG diphosphate after conversion to the triphosphate by nucleotide diphosphate kinase (3). There is no reported evidence that unphosphorylated 8-oxodG is a substrate for nucleotide salvage.…”

mentioning

confidence: 96%

“…1A). If not repaired, it can direct incorporation of dATP by DNA polymerases, resulting in G 3 T transversion mutations (1)(2)(3)(4). Likewise, 8-oxodG triphosphate (8-oxodGTP) is a mutagenic substrate for DNA synthesis because it can be incorporated opposite adenine and cytosine resulting in A 3 C and G 3 T transversions (5)(6)(7).…”

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confidence: 99%

“…To avoid mutations caused by 8-oxodG, Escherichia coli has evolved at least three enzymes that play pivotal roles in DNA repair (1)(2)(3). The DNA glycosylase formamidopyrimidine-DNA glycosylase (Fpg), also known as MutM, removes the oxidized base from 8-oxodG:dC base pairs.…”

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confidence: 99%

“…In the nucleotide pool, the pyrophosphohydrolase MutT hydrolyzes 8-oxodGTP to 8-oxodG monophosphate , reducing the concentration of 8-oxodGTP available for DNA synthesis. Mammals use repair enzymes that are functionally homologous to MutM, MutY, and MutT, including OGG1, MYH, and MutT homologous 1 (MTH1), respectively (3,8).…”

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Measurement of 7,8-dihydro-8-oxo-2′-deoxyguanosine metabolism in MCF-7 cells at low concentrations using accelerator mass spectrometry

Hah

Mundt

Kim

et al. 2007

Proc. Natl. Acad. Sci. U.S.A.

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Growing evidence suggests that oxidative damage to cells generates mutagenic 7,8-dihydro-8-oxo-2 -deoxyguanosine (8-oxodG), which may initiate diseases related to aging and carcinogenesis. Kinetic measurement of 8-oxodG metabolism and repair in cells has been hampered by poor assay sensitivity and by difficulty characterizing the flux of oxidized nucleotides through the relevant metabolic pathways. We report here the development of a sensitive and quantitative approach to characterizing the kinetics and metabolic sources of 8-oxodG in MCF-7 human breast cancer cells by accelerator mass spectrometry. We observed that [ 14 C]8-oxodG at medium concentrations of up to 2 pmol/ml was taken up by MCF-7 cells, phosphorylated to mono-, di-, and triphosphate derivatives, and incorporated into DNA. Oxidative stress caused by exposure of the cells to 17␤-estradiol resulted in a reduction in the rate of [ 14 C]8-oxodG incorporation into DNA and an increase in the ratio of 8-oxodG monophosphate (8-oxodGMP) to 8-oxodG triphosphate (8-oxodGTP) in the nucleotide pool. 17␤-Estradiolinduced oxidative stress up-regulated the nucleotide pool cleansing enzyme MTH1 and possibly other Nudix-related pyrophosphohydrolases. These data support the conclusion that 8-oxodGTP is formed in the nucleotide pool by both 8-oxodG metabolism and endogenous reactive oxygen species. The metabolism of 8-oxodG to 8-oxodGTP, followed by incorporation into DNA is a mechanism by which the cellular presence of this oxidized nucleoside can lead to mutations.DNA repair ͉ nucleoside metabolism ͉ oxidative stress ͉ breast cancer

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confidence: 96%

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confidence: 99%

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confidence: 99%

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Measurement of 7,8-dihydro-8-oxo-2′-deoxyguanosine metabolism in MCF-7 cells at low concentrations using accelerator mass spectrometry

Hah

Mundt

Kim

et al. 2007

Proc. Natl. Acad. Sci. U.S.A.

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“…Nucleotide pool sanitization is an important means by which organisms prevent the mutagenesis caused by damaged DNA precursors [6,12] …”

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Suppression of mutagenesis by 8-hydroxy-2′-deoxyguanosine 5′-triphosphate (7,8-dihydro-8-oxo-2′-deoxyguanosine 5′-triphosphate) by human MTH1, MTH2, and NUDT5

Hori

Satou

Harashima

et al. 2010

Free Radical Biology and Medicine

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To assess the functions of the three human MutT-type enzymes, MTH1, MTH2, and NUDT5, mutation induction by an oxidized form of dGTP, 8-hydroxy-2'-deoxyguanosine 5'-triphosphate (8-OH-dGTP, 7,8-dihydro-8-oxo-2'-deoxyguanosine 5'-triphosphate), was examined using human 293T cells treated with their specific siRNAs. Shuttle plasmid DNA containing the supF gene was first transfected into the cells, and then 8-OH-dGTP was introduced by means of osmotic pressure. Escherichia coli cells were transformed with the DNAs replicated in the treated cells.The knock-downs of the MTH1, MTH2, and NUDT5 proteins increased the A:TC:G substitution mutations induced by 8-OH-dGTP. In addition, the increase in the induced mutation frequency was more evident in the triple knock-down cells. These results indicate that all three of the human MTH1, MTH2, and NUDT5 proteins act as a defense against the mutagenesis induced by oxidized dGTP. 3Normal cellular metabolism produces endogenous reactive oxygen species (ROS). ROS are generated as byproducts of the mitochondrial electron transport chain, and certain cellular enzymes also generate ROS.Moreover, ROS are produced by environmental mutagens/carcinogens, including ionizing radiation and ultraviolet light. The formation of ROS leads to the oxidation of cellular components and disturbs their normal functions. The formation of oxidized DNA lesions is one of the causative factors of mutagenesis, carcinogenesis, neurodegeneration, and aging [1][2][3][4][5].DNA precursors (2'-deoxyribonucleotides) are also subjected to oxidative damage. The formation of oxidized DNA precursors is a potential source of mutagenesis [6]. 8-Hydroxy-2'-deoxyguanosine 5'-triphosphate (8-OH-dGTP, 7,8-dihydro-8-oxo-2'-deoxyguanosine 5'-triphosphate) is the major oxidation product of dGTP in in vitro oxidation reactions [7].8-OH-dGTP was reportedly present at a concentration of 1-10% relative to the unmodified dGTP in the mitochondrial nucleotide pool [8]. This oxidized form of dGTP is highly mutagenic in living cells when added exogenously [9][10][11] and is expected to act as an endogenous mutagen.Nucleotide pool sanitization is an important means by which organisms prevent the mutagenesis caused by damaged DNA precursors [6,12] Mutagenesis experiments293T cells (3 X 10 4 cells) were plated into 24-well dishes and were cultured in Dulbecco's modified Eagle medium supplemented with 10% fetal calf serum, at 37°C under a 5% CO 2 atmosphere for 24 hr. siRNAs (7.2 pmol each) were mixed with Lipofectamine (Invitrogen) and introduced into the cultured 293T cells according to the supplier's recommendations. In the triple knock-down experiment, an siRNA cocktail (MTH1; 3.6 pmol, MTH2; 7.2 pmol and NUDT5; 3.6 pmol) was used.After 24 Quantitative RT-PCR analysis of mRNATotal RNA was extracted from 293T cells using an RNeasy Mini Kit (Qiagen) combined with RNase-free DNase I (Takara, Otsu, Japan), for the degradation of the genomic DNA in the total RNA samples. First-strand cDNA synthesis was performed with 500 ng of t...

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Oxidative Stress in Adult Neurogenesis and in the Pathogenesis of Alzheimer Disease

Taupin

2011

Oxidative Stress in Vertebrates and Invertebrates

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Oxidative nucleotide damage: consequences and prevention

Cited by 203 publications

References 81 publications

Measurement of 7,8-dihydro-8-oxo-2′-deoxyguanosine metabolism in MCF-7 cells at low concentrations using accelerator mass spectrometry

Measurement of 7,8-dihydro-8-oxo-2′-deoxyguanosine metabolism in MCF-7 cells at low concentrations using accelerator mass spectrometry

Suppression of mutagenesis by 8-hydroxy-2′-deoxyguanosine 5′-triphosphate (7,8-dihydro-8-oxo-2′-deoxyguanosine 5′-triphosphate) by human MTH1, MTH2, and NUDT5

Oxidative Stress in Adult Neurogenesis and in the Pathogenesis of Alzheimer Disease

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