Prominent mitochondrial DNA recombination intermediates in human heart muscle

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Abstract
Recombination intermediates containing four‐way (Holliday) junctions are generated during DNA repair and replication in many systems, including yeast mitochondrial DNA (mtDNA). In contrast, convincing evidence for recombination in mammalian mtDNA is lacking. We have used two‐dimensional agarose‐gel electrophoresis to analyse non‐linear forms of mtDNA in human heart muscle. Replication intermediates from both the coupled and strand‐asynchronous mtDNA replication pathways were detected. An additional class of non‐linear molecules, with the electrophoretic properties of four‐way junctions, was also prominent. These molecules were insensitive to topoisomerase I or RNase H, but were diminished by branch migration or RuvC treatment. Junctional molecules were detected in all regions of the mitochondrial genome, were found in myocardial DNA from young and old adults, but were present at lower levels in skeletal muscle and placenta. We suggest that they could represent intermediates of mtDNA repair, given their prevalence in the oxyradical‐rich environment of heart muscle mitochondria.