Thermostable DNA polymerases can perform translesion synthesis using 8-oxoguanine and tetrahydrofuran-containing DNA templates

Biochim Biophys Acta. 2006 Jan;1764(1):97-104. doi: 10.1016/j.bbapap.2005.11.001. Epub 2005 Dec 1.

Abstract

The translesion synthesis (TLS) capacity of the thermostable DNA polymerases Taq, Tte and Tte-seq utilizing a synthetic abasic site, tetrahydrofuran (THF), and an 8-oxoguanine-containing DNA template was investigated. Measurements with human DNA polymerase beta were used as a "positive control". Thermostable DNA polymerases were observed to perform TLS with different specificities on both substrates. With a THF-containing template, dGMP was preferentially inserted by all the DNA polymerases. In the presence of Mn(II) as a cofactor, all the polymerases incorporated dCMP opposite 8-oxoguanine whereas, in the presence of Mg(II) ions, dAMP was incorporated. It was found that none of the thermophilic DNA polymerases utilized dTTP with either an 8-oxoguanine or a THF-containing template. In all cases, DNA duplex containing THF as damage was processed to full length less effectively than DNA duplex containing 8-oxoguanine.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Base Sequence
  • DNA / chemistry
  • DNA / genetics
  • DNA / metabolism
  • DNA Damage*
  • DNA Polymerase beta / metabolism
  • DNA Repair*
  • DNA Replication
  • DNA-Directed DNA Polymerase / chemistry
  • DNA-Directed DNA Polymerase / metabolism*
  • Deoxyribonucleotides / metabolism
  • Enzyme Stability
  • Furans / chemistry
  • Guanine / analogs & derivatives
  • Guanine / chemistry
  • Humans
  • In Vitro Techniques
  • Recombinant Proteins / chemistry
  • Recombinant Proteins / metabolism
  • Substrate Specificity
  • Taq Polymerase / chemistry
  • Taq Polymerase / metabolism
  • Thermus / enzymology
  • Thermus thermophilus / enzymology

Substances

  • Deoxyribonucleotides
  • Furans
  • Recombinant Proteins
  • tetrahydrofuran
  • 8-hydroxyguanine
  • Guanine
  • DNA
  • Taq Polymerase
  • DNA Polymerase beta
  • DNA-Directed DNA Polymerase