Hydroxylation of deoxyguanosine at 5' site of GG and GGG sequences in double-stranded DNA induced by carbamoyl radicals.
Free radicals generated by chemicals can cause sequence-specific DNA damage and play important roles in mutagenesis and carcinogenesis. Carbamoyl group (CONH2) and its derived groups (CONR2) occur as natural products and synthetic chemical compounds. We have investigated the DNA damage by carbamoyl radicals .(CONH2), one of carbon-centered radicals. Electron spin resonance (ESR) spectroscopic study has demonstrated that carbamoyl radicals were generated from formamide by treatment with H2O2 plus Cu(II), and from azodicarbonamide by treatment with Cu(II). We have investigated sequence specificity of DNA damage induced by carbamoyl radicals using 32P-labeled DNA fragments obtained from the human c-Ha-ras-1 and p53 genes. Treatment of double-stranded DNA with carbamoyl radicals induced an alteration of guanine residues, and subsequent treatment with piperidine or Fpg protein led to chain cleavages at 5'-G of GG and GGG sequences. Carbamoyl radicals enhanced Cu(II)/H2O2-mediated formation of 8-oxo-7,8-dihydro-2'-deoxyguanosine (8-oxodG) in double-stranded DNA more efficiently than that in single-stranded DNA. These results shows that carbamoyl radicals specifically induced hydroxylation of deoxyguanosine at 5' site of GG and GGG sequences in double-stranded DNA.
Department of Hygiene, Mie University School of Medicine, Edobashi 2-174, Tsu, Mie 514-8507, Japan.
Kaoru
K
Midorikawa
Kazutaka
K
Hirakawa
Shosuke
S
Kawanishi
D000080242:8-Hydroxy-2'-Deoxyguanosine; D001391:Azo Compounds / Q000737:chemistry / Q000378:metabolism / Q000494:pharmacology; D003300:Copper / Q000378:metabolism; D004247:DNA / Q000737:chemistry / Q000378:metabolism*; D004249:DNA Damage / Q000187:drug effects / Q000502:physiology; D003849:Deoxyguanosine / Q000031:analogs & derivatives* / Q000737:chemistry / Q000378:metabolism*; D005559:Formamides / Q000737:chemistry / Q000378:metabolism* / Q000494:pharmacology; D005609:Free Radicals / Q000737:chemistry / Q000378:metabolism*; D011905:Genes, ras; D006801:Humans; D006861:Hydrogen Peroxide / Q000378:metabolism / Q000494:pharmacology; D006900:Hydroxylation; D066298:In Vitro Techniques; D010618:Phenanthrolines / Q000494:pharmacology; D016159:Tumor Suppressor Protein p53 / Q000235:genetics
D001391:Azo Compounds; D005559:Formamides; D005609:Free Radicals; D010618:Phenanthrolines; D016159:Tumor Suppressor Protein p53; C031066:formamide; C004525:azodicarbonamide; D003300:Copper; D000080242:8-Hydroxy-2'-Deoxyguanosine; D004247:DNA; C002478:bathocuproine; D006861:Hydrogen Peroxide; D003849:Deoxyguanosine