DNA repair in plant mitochondria - a complete base excision repair pathway in potato tuber mitochondria

Physiol Plant. 2019 Jun;166(2):494-512. doi: 10.1111/ppl.12801. Epub 2018 Aug 31.

Abstract

Mitochondria are one of the major sites of reactive oxygen species (ROS) production in the plant cell. ROS can damage DNA, and this damage is in many organisms mainly repaired by the base excision repair (BER) pathway. We know very little about DNA repair in plants especially in the mitochondria. Combining proteomics, bioinformatics, western blot and enzyme assays, we here demonstrate that the complete BER pathway is found in mitochondria isolated from potato (Solanum tuberosum) tubers. The enzyme activities of three DNA glycosylases and an apurinic/apyrimidinic (AP) endonuclease (APE) were characterized with respect to Mg2+ dependence and, in the case of the APE, temperature sensitivity. Evidence for the presence of the DNA polymerase and the DNA ligase, which complete the repair pathway by replacing the excised base and closing the gap, was also obtained. We tested the effect of oxidative stress on the mitochondrial BER pathway by incubating potato tubers under hypoxia. Protein carbonylation increased significantly in hypoxic tuber mitochondria indicative of increased oxidative stress. The activity of two BER enzymes increased significantly in response to this oxidative stress consistent with the role of the BER pathway in the repair of oxidative damage to mitochondrial DNA.

MeSH terms

  • DNA Glycosylases / genetics
  • DNA Glycosylases / metabolism
  • DNA Repair / genetics*
  • DNA, Mitochondrial / genetics*
  • DNA, Plant / genetics*
  • DNA-(Apurinic or Apyrimidinic Site) Lyase / genetics
  • DNA-(Apurinic or Apyrimidinic Site) Lyase / metabolism
  • DNA-Directed DNA Polymerase / genetics
  • DNA-Directed DNA Polymerase / metabolism
  • Mitochondria / genetics
  • Mitochondria / metabolism
  • Reactive Oxygen Species / metabolism
  • Solanum tuberosum / genetics*
  • Solanum tuberosum / metabolism

Substances

  • DNA, Mitochondrial
  • DNA, Plant
  • Reactive Oxygen Species
  • DNA-Directed DNA Polymerase
  • DNA Glycosylases
  • DNA-(Apurinic or Apyrimidinic Site) Lyase