Journal
DNA REPAIR
Volume 11, Issue 8, Pages 649-656Publisher
ELSEVIER SCIENCE BV
DOI: 10.1016/j.dnarep.2012.05.004
Keywords
DNA replication; DNA polymerase epsilon; Ribonucleotides; Exonuclease; Proofreading
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Funding
- Division of Intramural Research of the National Institutes of Health (NIH) [Z01 ES065070]
- National Institute of Environmental Health Sciences (NIEHS)
- Swedish Research Council
- Swedish Cancer Society
- Smartafonden
- Basic Science-oriented Biotechnology, Medical Faculty of Umea University
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We have investigated the ability of the 3' exonuclease activity of Saccharomyces cerevisiae DNA polymerase epsilon (Pol epsilon) to proofread newly inserted ribonucleotides (rNMPs). During DNA synthesis in vitro, Pol epsilon proofreads ribonucleotides with apparent efficiencies that vary from none at some locations to more than 90% at others, with rA and rU being more efficiently proofread than rC and rG. Previous studies show that failure to repair ribonucleotides in the genome of rnh201 Delta strains that lack RNase H2 activity elevates the rate of short deletions in tandem repeat sequences. Here we show that this rate is increased by 2-4-fold in pol2-4 rnh2 Delta strains that are also defective in Pol e proofreading. In comparison, defective proofreading in these same strains increases the rate of base substitutions by more than 100-fold. Collectively, the results indicate that although proofreading of an 'incorrect' sugar is less efficient than is proofreading of an incorrect base, Pol epsilon does proofread newly inserted rNMPs to enhance genome stability. Published by Elsevier B.V.
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