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- W2000858063 abstract "DNA polymerases δ and ε (pol δ and ε) are the two major replicative polymerases in the budding yeast Saccharomyces cerevisiae. The fidelity of pol δ is influenced by its 3‘−5‘ proofreading exonuclease activity, which corrects misinsertion errors, and by enzyme cofactors. PCNA is a pol δ cofactor, called the sliding clamp, which increases the processivity of pol δ holoenzyme. This study measures the fidelity of 3‘−5‘ exonuclease-proficient and -deficient pol δ holoenzyme using a synthetic 30mer primer/100mer template in the presence and absence of PCNA. Although PCNA increases pol δ processivity, the presence of PCNA decreased pol δ fidelity 2−7-fold. In particular, wild-type pol δ demonstrated the following nucleotide substitution efficiencies for mismatches in the absence of PCNA: G·G, 0.728 × 10-4; T·G, 1.82 × 10-4; A·G, <0.01 × 10-4. In the presence of PCNA these values increased as follows: G·G, 1.30 × 10-4; T·G, 2.62 × 10-4; A·G, 0.074 × 10-4. A similar but smaller effect was observed for exonuclease-deficient pol δ (i.e., 2−4-fold increase in nucleotide substitution efficiencies in the presence of PCNA). Thus, the fidelity of wild-type pol δ in the presence of PCNA is more than 2 orders of magnitude lower than the fidelity of wild-type pol ε holoenzyme and is comparable to the fidelity of exonuclease-deficient pol ε holoenzyme." @default.
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- W2000858063 date "2003-11-12" @default.
- W2000858063 modified "2023-10-17" @default.
- W2000858063 title "Fidelity of DNA Polymerase δ Holoenzyme from <i>Saccharomyces cerevisiae</i>: The Sliding Clamp Proliferating Cell Nuclear Antigen Decreases Its Fidelity" @default.
- W2000858063 doi "https://doi.org/10.1021/bi0348359" @default.
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