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• W2909331505 endingPage "3207" @default.
• W2909331505 startingPage "3197" @default.
• W2909331505 abstract "4,6-Diamino-5-formamidopyrimidine (Fapy•dG) is an abundant form of oxidative DNA damage that is mutagenic and contributes to the pathogenesis of human disease. When Fapy•dG is in its nucleotide triphosphate form, Fapy•dGTP, it is inefficiently cleansed from the nucleotide pool by the responsible enzyme in Escherichia coli MutT and its mammalian homolog MTH1. Therefore, under oxidative stress conditions, Fapy•dGTP could become a pro-mutagenic substrate for insertion into the genome by DNA polymerases. Here, we evaluated insertion kinetics and high-resolution ternary complex crystal structures of a configurationally stable Fapy•dGTP analog, β-C-Fapy•dGTP, with DNA polymerase β. The crystallographic snapshots and kinetic data indicate that binding of β-C-Fapy•dGTP impedes enzyme closure, thus hindering insertion. The structures reveal that an active site residue, Asp276, positions β-C-Fapy•dGTP so that it distorts the geometry of critical catalytic atoms. Removal of this guardian side chain permits enzyme closure and increases the efficiency of β-C-Fapy•dG insertion opposite dC. These results highlight the stringent requirements necessary to achieve a closed DNA polymerase active site poised for efficient nucleotide incorporation and illustrate how DNA polymerase β has evolved to hinder Fapy•dGTP insertion." @default.
• W2909331505 created "2019-01-25" @default.
• W2909331505 creator A5004213173 @default.
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• W2909331505 creator A5082947137 @default.
• W2909331505 creator A5087642554 @default.
• W2909331505 date "2019-01-16" @default.
• W2909331505 modified "2023-10-16" @default.
• W2909331505 title "A guardian residue hinders insertion of a Fapy•dGTP analog by modulating the open-closed DNA polymerase transition" @default.
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• W2909331505 doi "https://doi.org/10.1093/nar/gkz002" @default.
• W2909331505 hasPubMedCentralId "https://www.ncbi.nlm.nih.gov/pmc/articles/6451102" @default.
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• W2909331505 hasPublicationYear "2019" @default.
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