Nucleotide pools dictate the identity and frequency of ribonucleotide incorporation in mitochondrial DNA.

Research paper by Anna-Karin AK Berglund, Clara C Navarrete, Martin K M MK Engqvist, Emily E Hoberg, Zsolt Z Szilagyi, Robert W RW Taylor, Claes M CM Gustafsson, Maria M Falkenberg, Anders R AR Clausen

Indexed on: 17 Feb '17Published on: 17 Feb '17Published in: PLoS genetics


Previous work has demonstrated the presence of ribonucleotides in human mitochondrial DNA (mtDNA) and in the present study we use a genome-wide approach to precisely map the location of these. We find that ribonucleotides are distributed evenly between the heavy- and light-strand of mtDNA. The relative levels of incorporated ribonucleotides reflect that DNA polymerase γ discriminates the four ribonucleotides differentially during DNA synthesis. The observed pattern is also dependent on the mitochondrial deoxyribonucleotide (dNTP) pools and disease-causing mutations that change these pools alter both the absolute and relative levels of incorporated ribonucleotides. Our analyses strongly suggest that DNA polymerase γ-dependent incorporation is the main source of ribonucleotides in mtDNA and argues against the existence of a mitochondrial ribonucleotide excision repair pathway in human cells. Furthermore, we clearly demonstrate that when dNTP pools are limiting, ribonucleotides serve as a source of building blocks to maintain DNA replication. Increased levels of embedded ribonucleotides in patient cells with disturbed nucleotide pools may contribute to a pathogenic mechanism that affects mtDNA stability and impair new rounds of mtDNA replication.