Replicative bypass studies of L-deoxyribonucleosides in Vitro and in E. coli cell

L-nucleosides were the most important antiviral lead compounds because they can inhibit viral DNA polymerase and DNA synthesis of many viruses, whereas they may lead to mutations in DNA replication and cause genomic instability. In this study, we reported the replicative bypass of L-deoxynucleosides in recombinant DNA by restriction enzyme-mediated assays to examine their impact on DNA replication in vitro and in E. coli cells. The results showed that a template L-dC inhibited Taq DNA polymerase reaction, whereas it can be bypassed by Vent (exo(-)) DNA polymerase as well as in cell replication, inserting correct nucleotides opposite L-dC. L-dG can be bypassed by Taq DNA polymerase and in E. coli cells, maintaining insertion of correct incoming nucleotides, and l-dG induced mutagenic replication by Vent (exo(-)) DNA polymerase. In contrast, L-dA can induced mutagenic replication in vitro and in E. coli cells. MD simulations were performed to investigate how DNA polymerase affected replicative bypass and mutations when D-nucleosides replaced with L-nucleosides. This study will provide a basis for the ability to assess the cytotoxic and mutagenic properties of the L-nucleoside drugs.

Automated summary

L-nucleosides have effects on DNA replication, and their bypass and mutagenic effects vary depending on the nucleoside. This study investigated the impact of L-deoxynucleosides on recombinant DNA replication using restriction enzyme-mediated assays, and performed molecular dynamics simulations to explore the effects of DNA polymerase. These findings provide insights into the cytotoxic and mutagenic properties of L-nucleoside drugs.