A mechanism for the exclusion of low-fidelity human Y-family DNA polymerases from base excision repair

Genes Dev. 2003 Nov 15;17(22):2777-85. doi: 10.1101/gad.1146103.

Abstract

The human Y-family DNA polymerases, Poliota, Poleta, and Polkappa, function in promoting replication through DNA lesions. However, because of their low fidelity, any involvement of these polymerases in DNA synthesis during base excision repair (BER) would be highly mutagenic. Mechanisms, therefore, must exist to exclude their participation in BER. Here, we show that although Poliota, Poleta, and Polkappa are all able to form a covalent Schiff base intermediate with the 5'-deoxyribose phosphate (5'-dRP) residue that results from the incision of DNA at an abasic site by an AP endonuclease, they all lack the ability for the subsequent catalytic removal of the 5'-dRP group. Instead, the covalent trapping of these polymerases by the 5'-dRP residue inhibits their DNA synthetic activity during BER. The unprecedented ability of these polymerases for robust Schiff base formation without the release of the 5'-dRP product provides a means of preventing their participation in the DNA synthetic step of BER, thereby avoiding the high incidence of mutagenesis and carcinogenesis that would otherwise occur.

Publication types

  • Comparative Study
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • DNA / metabolism*
  • DNA Primers / chemistry
  • DNA Repair / genetics*
  • DNA-Directed DNA Polymerase / genetics
  • DNA-Directed DNA Polymerase / isolation & purification
  • DNA-Directed DNA Polymerase / metabolism*
  • Exodeoxyribonucleases / pharmacology*
  • Glutathione Transferase / metabolism
  • Humans
  • Phosphorus-Oxygen Lyases / metabolism
  • Recombinant Fusion Proteins / genetics
  • Recombinant Fusion Proteins / isolation & purification
  • Recombinant Fusion Proteins / metabolism
  • Ribosemonophosphates / metabolism
  • Schiff Bases

Substances

  • DNA Primers
  • Recombinant Fusion Proteins
  • Ribosemonophosphates
  • Schiff Bases
  • 2-deoxyribose 5-phosphate
  • DNA
  • Glutathione Transferase
  • 5'-deoxyribose phosphate lyase
  • DNA polymerase iota
  • DNA-Directed DNA Polymerase
  • POLK protein, human
  • Rad30 protein
  • Exodeoxyribonucleases
  • Phosphorus-Oxygen Lyases