Cytotoxic and Mutagenic Properties of C3′-Epimeric Lesions of 2′-Deoxyribonucleosides in Escherichia coli Cells

5 years ago

Cytotoxic and Mutagenic Properties of C3′-Epimeric Lesions of 2′-Deoxyribonucleosides in Escherichia coli Cells

Nicholas J. Amato, Pengcheng Wang, Yinsheng Wang

Reactive oxygen species (ROS), resulting from endogenous metabolism and/or environmental exposure, can induce damage to the 2-deoxyribose moiety in DNA. Specifically, a hydrogen atom from each of the five carbon atoms in 2-deoxyribose can be abstracted by hydroxyl radical, and improper chemical repair of the ensuing radicals formed at the C1′, C3′, and C4′ positions can lead to the stereochemical inversion at these sites to yield epimeric 2-deoxyribose lesions. Although ROS-induced single-nucleobase lesions have been well studied, the biological consequences of the C3′-epimeric lesions of 2′-deoxynucleosides, i.e., 2′-deoxyxylonucleosides (dxN), have not been comprehensively investigated. Herein, we assessed the impact of dxN lesions on the efficiency and fidelity of DNA replication in Escherichia coli cells by conducting a competitive replication and adduct bypass assay with single-stranded M13 phage containing a site-specifically incorporated dxN. Our results revealed that, of the four dxN lesions, only dxG constituted a strong impediment to DNA replication, and intriguingly, dxT and dxC conferred replication bypass efficiencies higher than those of the unmodified counterparts. In addition, the three SOS-induced DNA polymerases (Pol II, Pol IV, and Pol V) did not play any appreciable role in bypassing these lesions. Among the four dxNs, only dxA directed a moderate frequency of dCMP misincorporation. These results provided important insights into the impact of the C3′-epimeric lesions on DNA replication in E. coli cells.

Publisher URL: http://dx.doi.org/10.1021/acs.biochem.7b00146

DOI: 10.1021/acs.biochem.7b00146