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Originally published In Press as doi:10.1074/jbc.M007867200 on November 2, 2000

J. Biol. Chem., Vol. 276, Issue 4, 2517-2522, January 26, 2001
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Translesion DNA Synthesis by Yeast DNA Polymerase eta  on Templates Containing N2-Guanine Adducts of 1,3-Butadiene Metabolites*

Irina G. Minko, M. Todd Washington, Louise Prakash, Satya Prakash, and R. Stephen LloydDagger

From the Sealy Center for Molecular Science, University of Texas Medical Branch, Galveston, Texas 77555-1071

Yeast DNA polymerase eta  can replicate through cis-syn cyclobutane pyrimidine dimers and 8-oxoguanine lesions with the same efficiency and accuracy as replication of an undamaged template. Previously, it has been shown that Escherichia coli DNA polymerases I, II, and III are incapable of bypassing DNA substrates containing N2-guanine adducts of stereoisomeric 1,3-butadiene metabolites. Here we showed that yeast polymerase eta  replicates DNA containing the monoadducts (S)-butadiene monoepoxide and (S,S)-butadiene diolepoxide N2-guanines albeit at an ~200-300-fold lower efficiency relative to the control guanine. Interestingly, nucleotide incorporation opposite the (R)-butadiene monoepoxide and the (R,R)-butadiene diolepoxide N2-guanines was ~10-fold less efficient than incorporation opposite their S stereoisomers. Polymerase eta  preferentially incorporates the correct nucleotide opposite and downstream of all four adducts, except that it shows high misincorporation frequencies for elongation of C paired with (R)-butadiene monoepoxide N2-guanine. Additionally, polymerase eta  does not bypass the (R,R)- and (S,S)-butadiene diolepoxide N2-guanine-N2-guanine intra- strand cross-links, and replication is completely blocked just prior to the lesion. Collectively, these data suggest that polymerase eta  can tolerate the geometric distortions in DNA conferred by the N2-guanine butadiene monoadducts but not the intrastrand cross-links.

* This work was supported in part by National Institutes of Health Grants ES05355, S11-ES10018, and ES06676 (to R. S. L.) and GM 19261 (to L. P.).The costs of publication of this article were defrayed in part by the payment of page charges. The article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

Dagger Holds the Mary Gibbs Jones Distinguished Chair in Environmental Toxicology from the Houston Endowment. To whom correspondence should be addressed: Sealy Center for Molecular Science, University of Texas Medical Branch, 5.144 MRB, 301 University Blvd., Galveston, TX 77555-1071; Tel.: 409-772-2179; Fax: 409-772-1790; E-mail: rslloyd@utmb.edu.

Copyright © 2001 by The American Society for Biochemistry and Molecular Biology, Inc.


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