ArticleBiochemistry2010
DNA polymerase fidelity: comparing direct competition of right and wrong dNTP substrates with steady state and pre-steady state kinetics.
Article in Biochemistry, 2010. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 22 papers.
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Who cites it
22 citing papers in PubMed.
- Understanding the Fidelity and Specificity of DNA Polymerase I.ACS omega · 2025Article
- A DNA Cleavage Assay Using Synthetic Oligonucleotide Containing a Single Site-Directed Lesion for In Vitro Base Excision Repair Study.Methods in molecular biology (Clifton, N.J.) · 2023Article
- Mechanisms of telomerase inhibition by oxidized and therapeutic dNTPs.Nature communications · 2020Article
- Promutagenicity of 8-Chloroguanine, A Major Inflammation-Induced Halogenated DNA Lesion.Molecules (Basel, Switzerland) · 2019Article
- DNA polymerase activity on synthetic N3'→P5' phosphoramidate DNA templates.Nucleic acids research · 2019Article
- High-resolution mapping of DNA polymerase fidelity using nucleotide imbalances and next-generation sequencing.Nucleic acids research · 2018Article
- Oxidative guanine base damage regulates human telomerase activity.Nature structural & molecular biology · 2016Article
- PCR: the price of infidelity.Nature methods · 2016Article
- Kinetic selection vs. free energy of DNA base pairing in control of polymerase fidelity.Proceedings of the National Academy of Sciences of the United States of America · 2016Article
- DNA Polymerases η and ζ Combine to Bypass O(2)-[4-(3-Pyridyl)-4-oxobutyl]thymine, a DNA Adduct Formed from Tobacco Carcinogens.Chemical research in toxicology · 2016Article
- Measuring specificity in multi-substrate/product systems as a tool to investigate selectivity in vivo.Biochimica et biophysica acta · 2016Review
- Determination of hepatitis delta virus ribozyme N(-1) nucleobase and functional group specificity using internal competition kinetics.Analytical biochemistry · 2015Article
- Determination of relative rate constants for in vitro RNA processing reactions by internal competition.Analytical biochemistry · 2014Article
- A genetic assay for transcription errors reveals multilayer control of RNA polymerase II fidelity.PLoS genetics · 2014Article
- A quantitative fluorescence-based steady-state assay of DNA polymerase.The FEBS journal · 2014Article
- Optimization of DNA recovery and amplification from non-carbonized archaeobotanical remains.PloS one · 2014Article
- Effect of β,γ-CHF- and β,γ-CHCl-dGTP halogen atom stereochemistry on the transition state of DNA polymerase β.Biochemistry · 2012Article
- Thermodynamic basis for the emergence of genomes during prebiotic evolution.PLoS computational biology · 2012Article
- Replication pauses of the wild-type and mutant mitochondrial DNA polymerase gamma: a simulation study.PLoS computational biology · 2011Article
- Effect of the Y955C mutation on mitochondrial DNA polymerase nucleotide incorporation efficiency and fidelity.Biochemistry · 2011Article
Corrections and comments
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Authors and funding
4 authors.
Funding
Abstract
DNA polymerase fidelity is defined as the ratio of right (R) to wrong (W) nucleotide incorporations when dRTP and dWTP substrates compete at equal concentrations for primer extension at the same site in the polymerase-primer-template DNA complex. Typically, R incorporation is favored over W by 10(3)-10(5)-fold, even in the absence of 3'-exonuclease proofreading. Straightforward in principle, a direct competition fidelity measurement is difficult to perform in practice because detection of a small amount of W is masked by a large amount of R. As an alternative, enzyme kinetics measurements to evaluate k(cat)/K(m) for R and W in separate reactions are widely used to measure polymerase fidelity indirectly, based on a steady state derivation by Fersht. A systematic comparison between direct competition and kinetics has not been made until now. By separating R and W products using electrophoresis, we have successfully taken accurate fidelity measurements for directly competing R and W dNTP substrates for 9 of the 12 natural base mispairs. We compare our direct competition results with steady state and pre-steady state kinetic measurements of fidelity at the same template site, using the proofreading-deficient mutant of Klenow fragment (KF(-)) DNA polymerase. All the data are in quantitative agreement.
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Registered trials
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