Abundant ribonucleotide incorporation into DNA by yeast replicative polymerases

Stephanie A. Nick McElhinny, Brian E. Watts, Dinesh Kumar, Danielle L. Watt, Else Britt Lundström, Peter M.J. Burgers, Erik Johansson, Andrei Chabes, Thomas A. Kunkel

Research output: Contribution to journalArticle

263 Scopus citations

Abstract

Measurements of nucleoside triphosphate levels in Saccharomyces cerevisiae reveal that the four rNTPs are in 36- to 190-fold molar excess over their corresponding dNTPs. During DNA synthesis in vitro using the physiological nucleoside triphosphate concentrations, yeast DNA polymerase ε, which is implicated in leading strand replication, incorporates one rNMP for every 1,250 dNMPs. Pol δ and Pol α, which conduct lagging strand replication, incorporate one rNMP for every 5,000 or 625 dNMPs, respectively. Discrimination against rNMP incorporation varies widely, in some cases by more than 100-fold, depending on the identity of the base and the template sequence context in which it is located. Given estimates of the amount of replication catalyzed by Pols α, δ, and ε, the results are consistent with the possibility that more than 10,000 rNMPs may be incorporated into the nuclear genome during each round of replication in yeast. Thus, rNMPs may be the most common noncanonical nucleotides introduced into the eukaryotic genome. Potential beneficial and negative consequences of abundant ribonucleotide incorporation into DNA are discussed, including the possibility that unrepaired rNMPs in DNA could be problematic because yeast DNA polymerase ε has difficulty bypassing a single rNMP present within a DNA template.

Original languageEnglish
Pages (from-to)4949-4954
Number of pages6
JournalProceedings of the National Academy of Sciences of the United States of America
Volume107
Issue number11
DOIs
StatePublished - Mar 16 2010

Keywords

  • DNA replication
  • Nucleotide precursors
  • Nucleotide selectivity

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    Nick McElhinny, S. A., Watts, B. E., Kumar, D., Watt, D. L., Lundström, E. B., Burgers, P. M. J., Johansson, E., Chabes, A., & Kunkel, T. A. (2010). Abundant ribonucleotide incorporation into DNA by yeast replicative polymerases. Proceedings of the National Academy of Sciences of the United States of America, 107(11), 4949-4954. https://doi.org/10.1073/pnas.0914857107