title: Nuclear Oxidation of a Major Peroxidation DNA Adduct, M1dG, in the Genome.
abstract: Chronic inflammation results in increased production of reactive oxygen species (ROS), which can oxidize cellular molecules including lipids and DNA. Our laboratory has shown that 3-(2-deoxy-β-d-erythro-pentofuranosyl)pyrimido[1,2-α]purin-10(3H)-one (M1dG) is the most abundant DNA adduct formed from the lipid peroxidation product, malondialdehyde, or the DNA peroxidation product, base propenal. M1dG is mutagenic in bacterial and mammalian cells and is repaired via the nucleotide excision repair system. Here, we report that M1dG levels in intact DNA were increased from basal levels of 1 adduct per 10(8) nucleotides to 2 adducts per 10(6) nucleotides following adenine propenal treatment of RKO, HEK293, or HepG2 cells. We also found that M1dG in genomic DNA was oxidized in a time-dependent fashion to a single product, 6-oxo-M1dG (to ∼ 5 adducts per 10(7) nucleotides), and that this oxidation correlated with a decline in M1dG levels. Investigations in RAW264.7 macrophages indicate the presence of high basal levels of M1dG (1 adduct per 10(6) nucleotides) and the endogenous formation of 6-oxo-M1dG. This is the first report of the production of 6-oxo-M1dG in genomic DNA in intact cells, and it has significant implications for understanding the role of inflammation in DNA damage, mutagenesis, and repair.
authors.affiliation: A.B. Hancock Jr. Memorial Laboratory for Cancer Research, Departments of †Biochemistry, ‡Chemistry, and §Pharmacology, Vanderbilt Institute of Chemical Biology, Center in Molecular Toxicology, Vanderbilt-Ingram Cancer Center, Vanderbilt University School of Medicine , Nashville, Tennessee 37232-0146, United States.
authors.forename: Orrette R
authors.initials: OR
authors.lastname: Wauchope
authors.affiliation: A.B. Hancock Jr. Memorial Laboratory for Cancer Research, Departments of †Biochemistry, ‡Chemistry, and §Pharmacology, Vanderbilt Institute of Chemical Biology, Center in Molecular Toxicology, Vanderbilt-Ingram Cancer Center, Vanderbilt University School of Medicine , Nashville, Tennessee 37232-0146, United States.
authors.forename: William N
authors.initials: WN
authors.lastname: Beavers
authors.affiliation: A.B. Hancock Jr. Memorial Laboratory for Cancer Research, Departments of †Biochemistry, ‡Chemistry, and §Pharmacology, Vanderbilt Institute of Chemical Biology, Center in Molecular Toxicology, Vanderbilt-Ingram Cancer Center, Vanderbilt University School of Medicine , Nashville, Tennessee 37232-0146, United States.
authors.forename: James J
authors.initials: JJ
authors.lastname: Galligan
authors.affiliation: A.B. Hancock Jr. Memorial Laboratory for Cancer Research, Departments of †Biochemistry, ‡Chemistry, and §Pharmacology, Vanderbilt Institute of Chemical Biology, Center in Molecular Toxicology, Vanderbilt-Ingram Cancer Center, Vanderbilt University School of Medicine , Nashville, Tennessee 37232-0146, United States.
authors.forename: Michelle M
authors.initials: MM
authors.lastname: Mitchener
authors.affiliation: A.B. Hancock Jr. Memorial Laboratory for Cancer Research, Departments of †Biochemistry, ‡Chemistry, and §Pharmacology, Vanderbilt Institute of Chemical Biology, Center in Molecular Toxicology, Vanderbilt-Ingram Cancer Center, Vanderbilt University School of Medicine , Nashville, Tennessee 37232-0146, United States.
authors.forename: Philip J
authors.initials: PJ
authors.lastname: Kingsley
authors.affiliation: A.B. Hancock Jr. Memorial Laboratory for Cancer Research, Departments of †Biochemistry, ‡Chemistry, and §Pharmacology, Vanderbilt Institute of Chemical Biology, Center in Molecular Toxicology, Vanderbilt-Ingram Cancer Center, Vanderbilt University School of Medicine , Nashville, Tennessee 37232-0146, United States.
authors.forename: Lawrence J
authors.initials: LJ
authors.lastname: Marnett
mesh_terms: D000225:Adenine / Q000031:analogs & derivatives / Q000633:toxicity; D000818:Animals; D002467:Cell Nucleus / Q000235:genetics; D002478:Cells, Cultured; D002853:Chromatography, Liquid; D018736:DNA Adducts / Q000737:chemistry*; D057809:HEK293 Cells; D006801:Humans; D015227:Lipid Peroxidation; D008264:Macrophages / Q000187:drug effects; D013058:Mass Spectrometry; D010084:Oxidation-Reduction; D011684:Purine Nucleosides / Q000737:chemistry*
chemical_list: C491625:3-(2'-deoxy-beta-D-erythro-pentofuranosyl)pyrimido(1,2-alpha)purin-10(3H)-one; D018736:DNA Adducts; D011684:Purine Nucleosides; C085912:9-(3-oxoprop-1-enyl)adenine; D000225:Adenine