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Nucleic Acids Research, 2002, Vol. 30, No. 7 1630-1638
© 2002 Oxford University Press

Response of human REV1 to different DNA damage: preferential dCMP insertion opposite the lesion

Yanbin Zhang, Xiaohua Wu, Olga Rechkoblit1, Nicholas E. Geacintov1, John-Stephen Taylor2 and Zhigang Wang*

306 Health Sciences Research Building, Graduate Center for Toxicology, University of Kentucky, Lexington, KY 40536, USA, 1Chemistry Department, New York University, New York, NY 10003, USA and 2Department of Chemistry, Washington University, St Louis, MO 63130, USA

REV1 functions in the DNA polymerase {zeta} mutagenesis pathway. To help understand the role of REV1 in lesion bypass, we have examined activities of purified human REV1 opposite various template bases and several different DNA lesions. Lacking a 3'->5' proofreading exonuclease activity, purified human REV1 exhibited a DNA polymerase activity on a repeating template G sequence, but catalyzed nucleotide insertion with 6-fold lower efficiency opposite a template A and 19–27-fold lower efficiency opposite a template T or C. Furthermore, dCMP insertion was greatly preferred regardless of the specific template base. Human REV1 inserted a dCMP efficiently opposite a template 8-oxoguanine, (+)-trans-anti-benzo[a]pyrene-N 2-dG, (–)-trans-anti-benzo[a]pyrene-N 2-dG and 1,N 6-ethenoadenine adducts, very inefficiently opposite an acetylaminofluorene-adducted guanine, but was unresponsive to a template TT dimer or TT (6–4) photoproduct. Surprisingly, the REV1 specificity of nucleotide insertion was very similar in response to different DNA lesions with greatly preferred C insertion and least frequent A insertion. By combining the dCMP insertion activity of human REV1 with the extension synthesis activity of human polymerase {kappa}, bypass of the trans-anti-benzo[a]pyrene-N 2 -dG adducts and the 1,N 6-ethenoadenine lesion was achieved by the two-polymerase two-step mechanism. These results suggest that human REV1 is a specialized DNA polymerase that may contribute to dCMP insertion opposite many types of DNA damage during lesion bypass.

* To whom correspondence should be addressed. Tel: +1 859 323 5784; Fax: +1 859 323 1059; Email: zwang{at}uky.edu


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