Translesion synthesis of abasic sites by yeast DNA polymerase epsilon

J Biol Chem. 2009 Nov 13;284(46):31555-63. doi: 10.1074/jbc.M109.043927. Epub 2009 Sep 23.

Abstract

Studies of replicative DNA polymerases have led to the generalization that abasic sites are strong blocks to DNA replication. Here we show that yeast replicative DNA polymerase epsilon bypasses a model abasic site with comparable efficiency to Pol eta and Dpo4, two translesion polymerases. DNA polymerase epsilon also exhibited high bypass efficiency with a natural abasic site on the template. Translesion synthesis primarily resulted in deletions. In cases where only a single nucleotide was inserted, dATP was the preferred nucleotide opposite the natural abasic site. In contrast to translesion polymerases, DNA polymerase epsilon with 3'-5' proofreading exonuclease activity bypasses only the model abasic site during processive synthesis and cannot reinitiate DNA synthesis. This characteristic may allow other pathways to rescue leading strand synthesis when stalled at an abasic site.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • DNA Damage*
  • DNA Polymerase II / chemistry*
  • DNA Polymerase II / genetics
  • DNA Polymerase II / metabolism*
  • DNA Polymerase beta / metabolism
  • DNA Primers / chemistry
  • DNA Replication*
  • DNA, Fungal / genetics*
  • DNA-Directed DNA Polymerase / metabolism
  • Escherichia coli Proteins / metabolism
  • Furans / chemistry
  • Glutathione Transferase / genetics
  • Glutathione Transferase / metabolism
  • Recombinant Fusion Proteins / genetics
  • Recombinant Fusion Proteins / isolation & purification
  • Recombinant Fusion Proteins / metabolism
  • Saccharomyces cerevisiae / enzymology*
  • Saccharomyces cerevisiae / genetics
  • Saccharomyces cerevisiae / growth & development

Substances

  • DNA Primers
  • DNA, Fungal
  • Escherichia coli Proteins
  • Furans
  • Recombinant Fusion Proteins
  • tetrahydrofuran
  • Glutathione Transferase
  • Dpo4 protein, E coli
  • DNA Polymerase II
  • DNA Polymerase beta
  • DNA-Directed DNA Polymerase
  • Rad30 protein