Unfilled gaps by polβ lead to aberrant ligation by LIG1 at the downstream steps of base excision repair pathway

Nucleic Acids Res. 2024 Apr 24;52(7):3810-3822. doi: 10.1093/nar/gkae104.

Abstract

Base excision repair (BER) involves the tightly coordinated function of DNA polymerase β (polβ) and DNA ligase I (LIG1) at the downstream steps. Our previous studies emphasize that defective substrate-product channeling, from gap filling by polβ to nick sealing by LIG1, can lead to interruptions in repair pathway coordination. Yet, the molecular determinants that dictate accurate BER remains largely unknown. Here, we demonstrate that a lack of gap filling by polβ leads to faulty repair events and the formation of deleterious DNA intermediates. We dissect how ribonucleotide challenge and cancer-associated mutations could adversely impact the ability of polβ to efficiently fill the one nucleotide gap repair intermediate which subsequently results in gap ligation by LIG1, leading to the formation of single-nucleotide deletion products. Moreover, we demonstrate that LIG1 is not capable of discriminating against nick DNA containing a 3'-ribonucleotide, regardless of base-pairing potential or damage. Finally, AP-Endonuclease 1 (APE1) shows distinct substrate specificity for the exonuclease removal of 3'-mismatched bases and ribonucleotides from nick repair intermediate. Overall, our results reveal that unfilled gaps result in impaired coordination between polβ and LIG1, defining a possible type of mutagenic event at the downstream steps where APE1 could provide a proofreading role to maintain BER efficiency.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • DNA / genetics
  • DNA / metabolism
  • DNA Damage
  • DNA Ligase ATP* / genetics
  • DNA Ligase ATP* / metabolism
  • DNA Ligases / genetics
  • DNA Ligases / metabolism
  • DNA Polymerase beta* / genetics
  • DNA Polymerase beta* / metabolism
  • DNA Repair*
  • DNA-(Apurinic or Apyrimidinic Site) Lyase / genetics
  • DNA-(Apurinic or Apyrimidinic Site) Lyase / metabolism
  • Excision Repair
  • Humans

Substances

  • DNA Polymerase beta
  • DNA Ligase ATP
  • LIG1 protein, human
  • DNA-(Apurinic or Apyrimidinic Site) Lyase
  • DNA
  • POLB protein, human
  • APEX1 protein, human
  • DNA Ligases