Evidence for double-strand break mediated mitochondrial DNA replication in Saccharomyces cerevisiae
| Autor: | Lynn Harrison, Rona S. Scott, Kanchanjunga Prasai, Lucy C. Robinson, Kelly Tatchell |
|---|---|
| Rok vydání: | 2017 |
| Předmět: |
DNA Replication
0301 basic medicine Saccharomyces cerevisiae Proteins Cell division DNA polymerase DNA polymerase II Genes Fungal Replication Origin Eukaryotic DNA replication Saccharomyces cerevisiae Genome Integrity Repair and Replication DNA Mitochondrial Models Biological 03 medical and health sciences 0302 clinical medicine Bacterial Proteins Control of chromosome duplication DNA-(Apurinic or Apyrimidinic Site) Lyase Genetics Humans DNA Breaks Double-Stranded DNA Fungal S phase biology DNA replication Cell biology DNA-Binding Proteins 030104 developmental biology Mutation MCF-7 Cells Mycobacterium marinum biology.protein 030217 neurology & neurosurgery Mitochondrial DNA replication |
| Zdroj: | Nucleic Acids Research |
| ISSN: | 1362-4962 0305-1048 |
| DOI: | 10.1093/nar/gkx443 |
| Popis: | The mechanism of mitochondrial DNA (mtDNA) replication in Saccharomyces cerevisiae is controversial. Evidence exists for double-strand break (DSB) mediated recombination-dependent replication at mitochondrial replication origin ori5 in hypersuppressive ρ− cells. However, it is not clear if this replication mode operates in ρ+ cells. To understand this, we targeted bacterial Ku (bKu), a DSB binding protein, to the mitochondria of ρ+ cells with the hypothesis that bKu would bind persistently to mtDNA DSBs, thereby preventing mtDNA replication or repair. Here, we show that mitochondrial-targeted bKu binds to ori5 and that inducible expression of bKu triggers petite formation preferentially in daughter cells. bKu expression also induces mtDNA depletion that eventually results in the formation of ρ0 cells. This data supports the idea that yeast mtDNA replication is initiated by a DSB and bKu inhibits mtDNA replication by binding to a DSB at ori5, preventing mtDNA segregation to daughter cells. Interestingly, we find that mitochondrial-targeted bKu does not decrease mtDNA content in human MCF7 cells. This finding is in agreement with the fact that human mtDNA replication, typically, is not initiated by a DSB. Therefore, this study provides evidence that DSB-mediated replication is the predominant form of mtDNA replication in ρ+ yeast cells. |
| Databáze: | OpenAIRE |
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