Published online 11 February 2004
Nucleic Acids Research, 2004, Vol. 32, No. 3 1122-1130
© 2004 Oxford University Press
Translesion synthesis of acetylaminofluorene-dG adducts by DNA polymerase 
is stimulated by yeast Rev1 protein
Graduate Center for Toxicology, University of Kentucky, Lexington, KY 40536, USA
*To whom correspondence should be addressed. Tel: +1 859 323 5784; Fax: +1 859 323 1059; Email: zwang{at}uky.edu
Translesion synthesis is an important mechanism in response to unrepaired DNA lesions during replication. The DNA polymerase 
(Pol) mutagenesis pathway is a major error-prone translesion synthesis mechanism requiring Pol 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 Pol was very inefficient in the bypass of the AAF-dG adduct. Combining Rev1 and Pol, however, led to a synergistic effect on translesion synthesis. Rev1 protein enhanced Pol-catalyzed nucleotide insertion opposite the AAF-dG adduct and strongly stimulated Pol-catalyzed extension from opposite the lesion. Rev1 also stimulated the deficient synthesis by Pol 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 Pol-catalyzed extension from opposite the AAF-dG adduct. These results suggest that translesion synthesis of AAF-dG adducts by Pol is stimulated by Rev1 protein in yeast. Consistent with the in vitro results, both Pol and Rev1 were found to be equally important for error-prone translesion synthesis across from AAF-dG DNA adducts in yeast cells.
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