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Papers In Press, published online ahead of print October 27, 2000
Sealy Center for Molecular Science, University of Texas Medical Branch, Galveston, TX 77555-1061
Corresponding Author: lprakash{at}scms.utmb.edu
DNA polymerase
J. Biol. Chem, 10.1074/jbc.M009049200
Submitted on October 3, 2000
Revised on October 27, 2000
Accepted on October 26, 2000
Mismatch extension ability of yeast and human DNA polymerase
(Pol) functions in error-free replication of UV damaged DNA, and in vitro, it efficiently bypasses a cis-syn thymine-thymine (T-T) dimer by incorporating two adenines opposite the lesion. Steady-state kinetic studies have shown that both yeast and human Pol are low fidelity enzymes, and they misincorporate nucleotides with a frequency of 10-2 to 10-3 on both undamaged and T-T dimer-containing DNA templates. To better understand the role of Pol in error-free translesion DNA synthesis, here we examine the ability of Pol to extend from base mismatches. We find that both yeast and human Pol extend from mismatched base pairs with a frequency of ~ 10-3 relative to matched base pairs. In the absence of efficient extension of mismatched primer termini, the ensuing dissociation of Pol from DNA may favor the excision of mismatched nucleotides by a proofreading exonuclease. Thus, we expect DNA synthesis by Pol to be more accurate than that predicted from the fidelity of nucleotide incorporation alone.
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