Nick sealing of polβ mismatch insertion products by LIG1 and LIG3α during 8-oxoG bypass leads to mutagenic or error-free base excision repair

Abstract

Base excision repair (BER) requires a coordination at the downstream steps involving gap filling by DNA polymerase (pol) β and subsequent nick sealing by DNA ligase (LIG) 1 or 3α. We previously reported that a failure in DNA ligase function, stemming from an impairment in nick sealing of polβ nucleotide insertion products, leads to faulty repair events. Yet, how the fidelity of 8-oxoG bypass by polβ affects the efficiency of ligation remains unclear. Here, we show that LIG1 and LIG3α seal the resulting nick repair product of polβ mutagenic insertion of dATP opposite 8-oxoG, while LIG3α exhibits an inability to ligate polβ dCTP:8-oxoG insertion product, demonstrating that the identity of BER ligase plays a critical role in repair outcomes at the final step. Furthermore, our results show that a lack of ribonucleotide insertion by polβ during 8-oxoG bypass diminishes the repair coordination with both ligases, highlighting the critical role of nucleotide selectivity in maintaining BER accuracy. Finally, our results reveal that AP-Endonuclease 1 (APE1) proofreads nick repair intermediates containing 3'-mismatches or ribonucleotides templating 8-oxoG. Overall, our findings provide a mechanistic insight into how the dual coding potential of the oxidative lesion in -anti versus -syn conformation could govern error-prone versus error-free repair outcomes, leading to deviations in the BER pathway coordination and the formation of deleterious DNA intermediates.

Keywords: 8oxoG; AP-Endonuclease 1; DNA ligase 1; DNA ligase 3; DNA polymerase β; base excision repair; oxidative DNA damage.