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M112146200v1
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Papers In Press, published online ahead of print February 15, 2002
J. Biol. Chem, 10.1074/jbc.M112146200
Submitted on December 19, 2001
Revised on February 12, 2002
Accepted on February 14, 2002

Yeast Rev1 protein is a G template specific DNA polymerase

Lajos Haracska, Satya Prakash, and Louise Prakash

Sealy Center for Molecular Science, University of Texas Medical Branch, Galveston, TX 77555-1061

Corresponding Author: lprakash{at}scms.utmb.edu

Rev1 protein of Saccharomyces cerevisiae functions with DNA polymerase zeta in mutagenic translesion synthesis. Because of the reported preferential incorporation of a C residue opposite an abasic site, Rev1 has been referred to as a deoxycytidyl transferase. Here, we use steady state kinetics to examine nucleotide incorporation by Rev1 opposite undamaged and damaged template residues. We show that Rev1 specifically inserts a C opposite template G, and it is ~25-, ~40-, and ~400-fold less efficient at inserting a C opposite an abasic site, an O6-methylguanine, and an 8-oxoguanine lesion, respectively. Rev1 misincorporates G, A, and T residues opposite template G with a frequency of ~10-3-10-4. Consistent with this, Rev1 replicates DNA containing a string of Gs in a template specific manner, but it has a low processivity incorporating 1.6 nucleotides per DNA binding event on the average. From these observations, we infer that Rev1 is a G template specific DNA polymerase.


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