title: Yeast Rev1 protein is a G template-specific DNA polymerase.
abstract: Rev1 protein of Saccharomyces cerevisiae functions with DNA polymerase zeta in mutagenic trans-lesion synthesis. Because of the reported preferential incorporation of a C residue opposite an abasic site, Rev1 has been referred to as a deoxycytidyltransferase. 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 residue opposite template G, and it is approximately 25-, 40-, and 400-fold less efficient at inserting a C residue opposite an abasic site, an O(6)-methylguanine, and an 8-oxoguanine lesion, respectively. Rev1 misincorporates G, A, and T residues opposite template G with a frequency of approximately 10(-3) to 10(-4). Consistent with this finding, 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.
authors.affiliation: Sealy Center for Molecular Science, University of Texas Medical Branch, Galveston, Texas 77555-1061, USA.
authors.forename: Lajos
authors.initials: L
authors.lastname: Haracska
authors.forename: Satya
authors.initials: S
authors.lastname: Prakash
authors.forename: Louise
authors.initials: L
authors.lastname: Prakash
mesh_terms: D001483:Base Sequence; D004249:DNA Damage; D017931:DNA Primers; D004251:DNA Transposable Elements; D004259:DNA-Directed DNA Polymerase / Q000378:metabolism*; D005656:Fungal Proteins / Q000378:metabolism*; D007700:Kinetics; D009713:Nucleotidyltransferases; D012441:Saccharomyces cerevisiae / Q000201:enzymology*; D029701:Saccharomyces cerevisiae Proteins; D013379:Substrate Specificity; D013698:Templates, Genetic; D000099270:Y-Family DNA Polymerases
chemical_list: D017931:DNA Primers; D004251:DNA Transposable Elements; D004259:DNA-Directed DNA Polymerase; D005656:Fungal Proteins; D009713:Nucleotidyltransferases; D029701:Saccharomyces cerevisiae Proteins; C425929:DNA polymerase zeta; C100951:REV1 protein, S cerevisiae; D000099270:Y-Family DNA Polymerases