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Title: The Impact of Minor-Groove N2-Alkyl-2'-deoxyguanosine Lesions on DNA Replication in Human Cells.

Authors: Wu, Jun; Du, Hua; Li, Lin; Price, Nathan E; Liu, Xiaochuan; Wang, Yinsheng

Published In ACS Chem Biol, (2019 08 16)

Abstract: Endogenous metabolites and exogenous chemicals can induce covalent modifications on DNA, producing DNA lesions. The N2 of guanine was shown to be a common alkylation site in DNA; however, not much is known about the influence of the size of the alkyl group in N2-alkyldG lesions on cellular DNA replication or how translesion synthesis (TLS) polymerases modulate DNA replication past these lesions in human cells. To answer these questions, we employ a robust shuttle vector method to investigate the impact of four N2-alkyldG lesions (i.e., with the alkyl group being a methyl, ethyl, n-propyl, or n-butyl group) on DNA replication in human cells. We find that replication through the N2-alkyldG lesions was highly efficient and accurate in HEK293T cells or isogenic CRISPR-engineered cells with deficiency in polymerase (Pol) ζ or Pol η. Genetic ablation of Pol ι, Pol κ, or Rev1, however, results in decreased bypass efficiencies and elicits substantial frequencies of G → A transition and G → T transversion mutations for these lesions. Moreover, further depletion of Pol ζ in Pol κ- or Pol ι-deficient cells gives rise to elevated rates of G → A and G → T mutations and substantially decreased bypass efficiencies. Cumulatively, we demonstrate that the error-free replication past the N2-alkyldG lesions is facilitated by a specific subset of TLS polymerases, and we find that longer alkyl chains in these lesions induce diminished bypass efficiency and fidelity in DNA replication.

PubMed ID: 31347832 Exiting the NIEHS site

MeSH Terms: Alkylation; DNA Damage; DNA Repair; DNA Replication/drug effects*; DNA-Directed DNA Polymerase/physiology; DNA/genetics; DNA/metabolism*; Deoxyguanosine/analogs & derivatives*; Deoxyguanosine/metabolism*; Deoxyguanosine/toxicity; HEK293 Cells; Humans; Molecular Structure; Mutation; Nucleic Acid Conformation

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