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Ribonucleotides incorporated by the yeast mitochondrial DNA polymerase are not repaired

Journal article
Authors P. H. Wanrooij
Martin K M Engqvist
J. M. E. Forslund
Clara Navarrete
A. K. Nilsson
J. Sedman
S. Wanrooij
Anders R Clausen
A. Chabes
Published in Proceedings of the National Academy of Sciences of the United States of America
Volume 114
Issue 47
Pages 12466-12471
ISSN 0027-8424
Publication year 2017
Published at Institute of Biomedicine
Institute of Biomedicine, Department of Medical Biochemistry and Cell Biology
Pages 12466-12471
Language en
Links dx.doi.org/10.1073/pnas.1713085114
Keywords DNA replication, DNTP, Mitochondrial DNA, Ribonucleotide excision repair, Ribonucleotide incorporation, cell nucleus DNA, deoxyribonucleoside triphosphate, ribonucleotide, Article, biochemical analysis, excision repair, mitochondrial genome, nonhuman, priority journal, yeast
Subject categories Cell and molecular biology, Cell and Molecular Biology

Abstract

Incorporation of ribonucleotides into DNA during genome replication is a significant source of genomic instability. The frequency of ribonucleotides in DNA is determined by deoxyribonucleoside triphosphate/ribonucleoside triphosphate (dNTP/rNTP) ratios, by the ability of DNA polymerases to discriminate against ribonucleotides, and by the capacity of repair mechanisms to remove incorporated ribonucleotides. To simultaneously compare how the nuclear and mitochondrial genomes incorporate and remove ribonucleotides, we challenged these processes by changing the balance of cellular dNTPs. Using a collection of yeast strains with altered dNTP pools, we discovered an inverse relationship between the concentration of individual dNTPs and the amount of the corresponding ribonucleotides incorporated in mitochondrial DNA, while in nuclear DNA the ribonucleotide pattern was only altered in the absence of ribonucleotide excision repair. Our analysis uncovers major differences in ribonucleotide repair between the two genomes and provides concrete evidence that yeast mitochondria lack mechanisms for removal of ribonucleotides incorporated by the mtDNA polymerase. Furthermore, as cytosolic dNTP pool imbalances were transmitted equally well into the nucleus and the mitochondria, our results support a view of the cytosolic and mitochondrial dNTP pools in frequent exchange. © 2017, National Academy of Sciences. All rights reserved.

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