Miscoding events during DNA synthesis past the nitration-damaged base 8-nitroguanine

Laboratory of Chemical Biology, Department of Pharmacological Sciences, State University of New York at Stony Brook, Stony Brook, New York 11794-8651, USA.
Biochemistry (Impact Factor: 3.01). 07/2005; 44(25):9238-45. DOI: 10.1021/bi050276p
Source: PubMed

ABSTRACT 8-Nitro-2'-deoxyguanosine (8-NO(2)-dG) DNA adducts are induced by the reactive nitrogen species and may be associated with the development of cancer in inflammatory tissues. To explore the miscoding potential of 8-NO(2)-dG adduct, an oligodeoxynucleotide containing a single 8-NO(2)-dG adduct was prepared by photochemical synthesis and used as a template in primer extension reactions catalyzed by mammalian DNA polymerases (pol). Primer extension reactions catalyzed by pol alpha or beta were strongly retarded at the 8-NO(2)-dG lesion; a fraction of primers was extended past the lesion by incorporating preferentially dCMP, the correct base, opposite the lesion, accompanied by lesser amounts of dAMP and dGMP incorporation. In contrast, primer extension reactions catalyzed by pol eta or a truncated form of pol kappa (pol kappaDeltaC) readily extended past the 8-NO(2)-dG lesion. Pol eta and kappaDeltaC showed more broad miscoding spectra; direct incorporations of dCMP and dAMP were observed, along with lesser amounts of dGMP and dTMP incorporations and deletions. The miscoding frequencies induced by pol eta and kappaDeltaC were at least 8 times higher than that of pol alpha or beta. Miscoding frequency and specificity of 8-NO(2)-dG varied depending on the DNA polymerases used. These observations were supported by steady-state kinetic studies. 8-NO(2)-dG adduct may play an important role in initiating inflammation driven carcinogenesis.

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    • "Cells deficient in Rev1 and Rev3, subunits of DNA polymerase í µí¼, were hypersensitive to nitrative stress, and translesion DNA synthesis past apurinic sites mediated by this polymerase might contribute to extensive point mutations [55]. It has been reported that adenine is preferentially incorporated opposite 8-nitroguanine during DNA synthesis catalyzed by polymerase í µí¼‚ and í µí¼…ΔC in a cell-free system, suggesting that G:C to T:A transversions can occur [56]. In the ONOO − -treated supF shuttle vector, which was replicated in host Escherichia coli cells, the majority of mutations occurred at G:C base pairs, predominantly involving G:C to T:A transversions [57]. "
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    Oxidative Medicine and Cellular Longevity 12/2013; 2013:387014. DOI:10.1155/2013/387014 · 3.36 Impact Factor
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    • "Recently, 8-nitroguanosine has been reported to be a highly redox-active molecule (Sawa et al., 2003; Zaki et al., 2005). More importantly, experimental evidence has suggested that 8-nitroguanine is a mutagenic DNA lesion, which preferentially leads to G™T transversions (Yermilov et al., 1995; Suzuki et al., 2005), in addition to 8-oxodG (Shibutani et al., 1991; Bruner et al., 2000). Indeed, G™T transversions have been observed in vivo in the ras gene (Bos, 1988) and the p53 tumor suppressor gene in lung and liver cancer (Takahashi et al., 1989; Hsu et al., 1991). "
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