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National Institute of Environmental Health Sciences

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Your Environment. Your Health.

Publication Detail

Title: Basis of miscoding of the DNA adduct N2,3-ethenoguanine by human Y-family DNA polymerases.

Authors: Zhao, Linlin; Pence, Matthew G; Christov, Plamen P; Wawrzak, Zdzislaw; Choi, Jeong-Yun; Rizzo, Carmelo J; Egli, Martin; Guengerich, F Peter

Published In J Biol Chem, (2012 Oct 12)

Abstract: N(2),3-Ethenoguanine (N(2),3-εG) is one of the exocyclic DNA adducts produced by endogenous processes (e.g. lipid peroxidation) and exposure to bioactivated vinyl monomers such as vinyl chloride, which is a known human carcinogen. Existing studies exploring the miscoding potential of this lesion are quite indirect because of the lability of the glycosidic bond. We utilized a 2'-fluoro isostere approach to stabilize this lesion and synthesized oligonucleotides containing 2'-fluoro-N(2),3-ε-2'-deoxyarabinoguanosine to investigate the miscoding potential of N(2),3-εG by Y-family human DNA polymerases (pols). In primer extension assays, pol η and pol κ replicated through N(2),3-εG, whereas pol ι and REV1 yielded only 1-base incorporation. Steady-state kinetics revealed that dCTP incorporation is preferred opposite N(2),3-εG with relative efficiencies in the order of pol κ > REV1 > pol η ≈ pol ι, and dTTP misincorporation is the major miscoding event by all four Y-family human DNA pols. Pol ι had the highest dTTP misincorporation frequency (0.71) followed by pol η (0.63). REV1 misincorporated dTTP and dGTP with much lower frequencies. Crystal structures of pol ι with N(2),3-εG paired to dCTP and dTTP revealed Hoogsteen-like base pairing mechanisms. Two hydrogen bonds were observed in the N(2),3-εG:dCTP base pair, whereas only one appears to be present in the case of the N(2),3-εG:dTTP pair. Base pairing mechanisms derived from the crystal structures explain the slightly favored dCTP insertion for pol ι in steady-state kinetic analysis. Taken together, these results provide a basis for the mutagenic potential of N(2),3-εG.

PubMed ID: 22910910 Exiting the NIEHS site

MeSH Terms: Cell-Free System/chemistry; Cell-Free System/metabolism; DNA Adducts/chemistry*; DNA Adducts/genetics; DNA Adducts/metabolism; DNA Polymerase beta/chemistry*; DNA Polymerase beta/genetics; DNA Polymerase beta/metabolism; DNA-Directed DNA Polymerase/chemistry*; DNA-Directed DNA Polymerase/genetics; DNA-Directed DNA Polymerase/metabolism; Guanine/analogs & derivatives*; Guanine/chemistry; Guanine/metabolism; Humans; Nuclear Proteins/chemistry*; Nuclear Proteins/genetics; Nuclear Proteins/metabolism; Nucleotidyltransferases/chemistry*; Nucleotidyltransferases/genetics; Nucleotidyltransferases/metabolism

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