Effects of 5′,8-Cyclodeoxyadenosine Triphosphates on DNA Synthesis

Hydroxyl radicals generate a broad range of DNA lesions in living cells. Cyclopurine deoxynucleosides (CPUs) are a biologically significant class of oxidative DNA lesions due to their helical distortion and chemically stability. The CPUs on DNA are substrates for the nucleotide excision repair (NER) but not for base excision repair or direct damage reversal. Moreover, these lesions block DNA and RNA polymerases, resulting in cell death. Here, we describe the chemical synthesis of 5′<i>S</i> and 5′<i>R</i> isomers of 5′,8-cyclodeoxyadenosine triphosphate (cdATP) and demonstrate their ability to be incorporated into DNA by replicative DNA polymerases. DNA synthesis assays revealed that the incorporation of the stereoisomeric cdATPs strongly inhibits DNA polymerase reactions. Surprisingly, the two stereoisomers had different mutagenic profiles, since the <i>S</i> isomer of cdATP could be inserted opposite to the dTMP, but the <i>R</i> isomer of cdATP could be inserted opposite to the dCMP. Kinetic analysis revealed that the <i>S</i> isomer of cdATP could be incorporated more efficiently (25.6 μM^–1 min^–1) than the <i>R</i> isomer (1.13 μM^–1 min^–1) during DNA synthesis. Previous data showed that the <i>S</i> isomer in DNA blocked DNA synthesis and the exonuclease activity of DNA polymerase and is less efficiently repaired by NER. This indicates that the <i>S</i> isomer has a tendency to accumulate on the genome DNA, and as such, the <i>S</i> isomer of cdATP may be a candidate cytotoxic drug