Protein-template-directed synthesis across an acrolein-derived DNA adduct by yeast Rev1 DNA polymerase

Structure. 2008 Feb;16(2):239-45. doi: 10.1016/j.str.2007.12.009.

Abstract

Acrolein is generated as the end product of lipid peroxidation and is also a ubiquitous environmental pollutant. Its reaction with the N2 of guanine leads to a cyclic gamma-HOPdG adduct that presents a block to normal replication. We show here that yeast Rev1 incorporates the correct nucleotide C opposite a permanently ring-closed form of gamma-HOPdG (PdG) with nearly the same efficiency as opposite an undamaged G. The structural basis of this action lies in the eviction of the PdG adduct from the Rev1 active site, and the pairing of incoming dCTP with a "surrogate" arginine residue. We also show that yeast Polzeta can carry out the subsequent extension reaction. Together, our studies reveal how the exocyclic PdG adduct is accommodated in a DNA polymerase active site, and they show that the combined action of Rev1 and Polzeta provides for accurate and efficient synthesis through this potentially carcinogenic DNA lesion.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • Acrolein / chemistry
  • Arginine / chemistry
  • Base Pairing
  • Binding Sites
  • Crystallography, X-Ray
  • DNA / biosynthesis
  • DNA / chemistry
  • DNA Adducts / chemistry*
  • DNA Damage
  • DNA-Directed DNA Polymerase / metabolism
  • Deoxycytosine Nucleotides / chemistry
  • Deoxycytosine Nucleotides / metabolism
  • Environmental Pollutants / chemistry
  • Models, Molecular
  • Nucleotidyltransferases / chemistry*
  • Nucleotidyltransferases / metabolism
  • Saccharomyces cerevisiae Proteins / chemistry*
  • Saccharomyces cerevisiae Proteins / metabolism
  • Templates, Genetic

Substances

  • DNA Adducts
  • Deoxycytosine Nucleotides
  • Environmental Pollutants
  • Saccharomyces cerevisiae Proteins
  • 2'-deoxycytidine 5'-triphosphate
  • Acrolein
  • DNA
  • Arginine
  • DNA polymerase zeta
  • Nucleotidyltransferases
  • REV1 protein, S cerevisiae
  • DNA-Directed DNA Polymerase

Associated data

  • PDB/3BJY