Prominent mitochondrial DNA recombination intermediates in human heart muscle

Analysis of heart muscle mitochondrial DNA in the present study reveals the presence, in addition to these replication intermediate, of prevalent recombination intermediates

Olli A Kajander


Scholarcy highlights

  • They can be visualized by two-dimensional neutral–neutral agarose-gel electrophoresis as a so-called X-spike, located at double unit-length for the fragment being analysed
  • The strand-specific nucleases-resistant species from this fragment were similar to those seen previously in human placenta or cells recovering from mitochondrial DNA depletion, with a more prominent, complete Y-arc
  • As in cells recovering from mtDNA depletion, the major SSN-sensitive species from the orthodox mode replication-containing fragment formed a diffuse arc below the track of the standard Y-arc, ending near to a prominent spot
  • Human heart mtDNA includes a prominent class of non-linear molecules, with the properties of recombination junctions
  • The X-spike closely resembles those which accumulate in yeast mtDNA when resolution of four-way junctions is blocked by deletion of the CCE1 gene
  • This is the first clear demonstration of recombination intermediates in mammalian mtDNA. Their abundance varies between tissues, being high in heart, intermediate in skeletal muscle or placenta, but low or absent in
  • To promote branch migration of possible Holliday junction structures, gel slabs from the first-dimension electrophoresis were incubated for 4 h at 65°C in 10 mM Tris–HCl, 0.1 M NaCl, 0.1 mM EDTA pH 8.0

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