Translesion synthesis of acetylaminofluorene-dG adducts by DNA polymerase zeta is stimulated by yeast Rev1 protein

Nucleic Acids Res. 2004 Feb 11;32(3):1122-30. doi: 10.1093/nar/gkh279. Print 2004.


Translesion synthesis is an important mechanism in response to unrepaired DNA lesions during replication. The DNA polymerase zeta (Polzeta) mutagenesis pathway is a major error-prone translesion synthesis mechanism requiring Polzeta and Rev1. In addition to its dCMP transferase, a non-catalytic function of Rev1 is suspected in cellular response to certain types of DNA lesions. However, it is not well understood about the non-catalytic function of Rev1 in translesion synthesis. We have analyzed the role of Rev1 in translesion synthesis of an acetylaminofluorene (AAF)-dG DNA adduct. Purified yeast Rev1 was essentially unresponsive to a template AAF-dG DNA adduct, in contrast to its efficient C insertion opposite a template 1,N6-ethenoadenine adduct. Purified yeast Polzeta was very inefficient in the bypass of the AAF-dG adduct. Combining Rev1 and Polzeta, however, led to a synergistic effect on translesion synthesis. Rev1 protein enhanced Polzeta-catalyzed nucleotide insertion opposite the AAF-dG adduct and strongly stimulated Polzeta-catalyzed extension from opposite the lesion. Rev1 also stimulated the deficient synthesis by Polzeta at the very end of undamaged DNA templates. Deleting the C-terminal 205 aa of Rev1 did not affect its dCMP transferase activity, but abolished its stimulatory activity on Polzeta-catalyzed extension from opposite the AAF-dG adduct. These results suggest that translesion synthesis of AAF-dG adducts by Polzeta is stimulated by Rev1 protein in yeast. Consistent with the in vitro results, both Polzeta and Rev1 were found to be equally important for error-prone translesion synthesis across from AAF-dG DNA adducts in yeast cells.

Publication types

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

MeSH terms

  • 2-Acetylaminofluorene / analogs & derivatives*
  • 2-Acetylaminofluorene / metabolism*
  • Base Sequence
  • DNA / biosynthesis*
  • DNA Adducts / biosynthesis
  • DNA Adducts / metabolism*
  • DNA Damage*
  • DNA-Directed DNA Polymerase / metabolism*
  • DNA-Directed DNA Polymerase / physiology
  • Deoxycytidine Monophosphate / metabolism
  • Deoxyguanosine / analogs & derivatives*
  • Deoxyguanosine / biosynthesis
  • Deoxyguanosine / metabolism*
  • Mutagenesis
  • Nucleotidyltransferases / metabolism
  • Nucleotidyltransferases / physiology*
  • Saccharomyces cerevisiae Proteins / metabolism
  • Saccharomyces cerevisiae Proteins / physiology*
  • Templates, Genetic
  • Yeasts / enzymology
  • Yeasts / genetics
  • Yeasts / metabolism


  • DNA Adducts
  • Saccharomyces cerevisiae Proteins
  • Deoxycytidine Monophosphate
  • N-(deoxyguanosin-8-yl)acetylaminofluorene
  • DNA
  • 2-Acetylaminofluorene
  • DNA polymerase zeta
  • Nucleotidyltransferases
  • REV1 protein, S cerevisiae
  • DNA-Directed DNA Polymerase
  • Deoxyguanosine