Inosine is a key intermediate in de novo purine nucleotide biosynthesis in cells. Inosine is known to be mutagenic when it is present in DNA, in place of adenine via deamination, by facilitating the incorporation of dCTP exclusively, resulting in A:T to G:C mutation. The structural basis for the mutagenicity of inosine bypass has been reported in some DNA polymerases including human DNA polymerase eta (polη). However, the structural and biochemical basis for the mutagenic potential of the incorporation of deoxyinosine triphosphate (dITP) into DNA remains poorly understood. To gain insights into the mutagenic potential of the incorporation of inosine into DNA, we conducted structural and kinetic studies of human polη incorporating dITP across undamaged DNA template containing dC or dT. Polη incorporated dITP opposite dC 14-fold more efficiently than opposite dT, indicating that dITP incorporation by polη can be mutagenic unlike the bypass of inosine by polη, which incorporated dCTP almost exclusively opposite the templating inosine over dTTP (70:1). Polη-dC:dITP crystal structure showed that the incoming dITP formed Watson-Crick base pair along with wobble base pair via 4-imino-2-keto tautomer of cytosine diminishing the catalytic efficiency compared to dGTP incorporation across dC. In addition, the crystal structure of polη-dT:dITP revealed that dT and dITP formed Watson-Crick like base pair via 4-enol-2-keto tautomer of thymine, reinforced by wobble base pair via 4-keto-2-keto tautomer of thymine resulting in the increased mutagenicity of dITP incorporation (14:1 across dC and dT), which is 14-fold higher than dGTP incorporation by polη (190:1 across dC and dT).
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Cover Image
Cover Image
The cover image is a fluorescent microscopy image showing colocalization of HA-GFP -NPT2A Arg495Cys (green) with FLAG-NHERF1 (red) in OK cells. Treatment with PTH does not change NPT2A Arg495Cys-NHERF1 colocalization at the apical membrane. For more information see the article by Sneddon and colleagues (pp. 685–699) in this issue. The image was provided by Tatyana Mamonova.
Mutagenic incorporation of inosine into DNA via T:I mismatch formation by human DNA polymerase eta (polη)
Jameson R. Averill, Hunmin Jung; Mutagenic incorporation of inosine into DNA via T:I mismatch formation by human DNA polymerase eta (polη). Biochem J 17 May 2023; 480 (9): 649–664. doi: https://doi.org/10.1042/BCJ20230029
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