Mitochondrial tyrosyl-DNA phosphodiesterase 2 and its TDP2S short isoform

EMBO Rep. 2018 Mar;19(3):e42139. doi: 10.15252/embr.201642139. Epub 2018 Feb 9.

Abstract

Tyrosyl-DNA phosphodiesterase 2 (TDP2) repairs abortive topoisomerase II cleavage complexes. Here, we identify a novel short isoform of TDP2 (TDP2S) expressed from an alternative transcription start site. TDP2S contains a mitochondrial targeting sequence, contributing to its enrichment in the mitochondria and cytosol, while full-length TDP2 contains a nuclear localization signal and the ubiquitin-associated domain in the N-terminus. Our study reveals that both TDP2 isoforms are present and active in the mitochondria. Comparison of isogenic wild-type (WT) and TDP2 knockout (TDP2-/-/-) DT40 cells shows that TDP2-/-/- cells are hypersensitive to mitochondrial-targeted doxorubicin (mtDox), and that complementing TDP2-/-/- cells with human TDP2 restores resistance to mtDox. Furthermore, mtDox selectively depletes mitochondrial DNA in TDP2-/-/- cells. Using CRISPR-engineered human cells expressing only the TDP2S isoform, we show that TDP2S also protects human cells against mtDox. Finally, lack of TDP2 in the mitochondria reduces the mitochondria transcription levels in two different human cell lines. In addition to identifying a novel TDP2S isoform, our report demonstrates the presence and importance of both TDP2 isoforms in the mitochondria.

Keywords: DNA repair; mitochondria; mitochondrial DNA; topoisomerase.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, N.I.H., Intramural

MeSH terms

  • Alternative Splicing / genetics
  • Cell Line, Tumor
  • DNA-Binding Proteins
  • Doxorubicin / pharmacology*
  • Drug Resistance, Neoplasm / genetics*
  • Gene Expression Regulation, Enzymologic / drug effects
  • Gene Knockout Techniques
  • Humans
  • Mitochondria / drug effects
  • Mitochondria / genetics
  • Neoplasms / drug therapy*
  • Neoplasms / genetics
  • Neoplasms / pathology
  • Nuclear Proteins / antagonists & inhibitors
  • Nuclear Proteins / genetics*
  • Phosphoric Diester Hydrolases
  • Protein Isoforms / genetics
  • Transcription Factors / antagonists & inhibitors
  • Transcription Factors / genetics*

Substances

  • DNA-Binding Proteins
  • Nuclear Proteins
  • Protein Isoforms
  • Transcription Factors
  • Doxorubicin
  • Phosphoric Diester Hydrolases
  • TDP2 protein, human