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Transcript availability dictates the balance between strand-asynchronous and strand-coupled mitochondrial DNA

Tricia J Cluett1, Gokhan Akman2, Aurelio Reyes1

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Mitochondrial DNA replication mechanisms are influenced by mitochondrial transcripts. Increased Twinkle DNA helicase expression shifts replication to a coupled mechanism, correlating with lower transcript levels, supporting transcripts

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Area of Science:

  • Molecular Biology
  • Genetics
  • Cell Biology

Background:

  • Mammalian mitochondria utilize diverse DNA replication strategies, with strand-asynchronous synthesis often prevailing over coupled leading/lagging-strand synthesis.
  • Factors governing these distinct replication mechanisms and the role of transient transcript incorporation (bootlaces) in strand-asynchronous replication remain poorly understood.

Purpose of the Study:

  • To investigate the influence of Twinkle DNA helicase expression on mitochondrial DNA replication mechanisms.
  • To determine the correlation between mitochondrial transcript levels and the predominant mode of DNA replication.

Main Methods:

  • Elevated expression of Twinkle DNA helicase in human mitochondria.
  • Analysis of DNA replication mechanisms (strand-asynchronous vs. coupled).
  • Quantification of mitochondrial transcript levels.

Main Results:

  • Increased Twinkle DNA helicase induced bidirectional, coupled leading and lagging-strand DNA synthesis, reducing strand-asynchronous replication.
  • This switch correlated with decreased steady-state levels of specific mitochondrial transcripts.
  • Strand-asynchronous replication persisted in the mitochondrial DNA minor arc where transcript levels remained high.

Conclusions:

  • Mitochondrial transcripts play a critical role in mediating the strand-asynchronous mechanism of mitochondrial DNA replication.
  • Mitochondrial RNA availability and subsequent RNA/DNA hybrid formation are potential regulators of mitochondrial DNA replication mechanisms.