Probing the Telomere Damage Response

Methods Mol Biol. 2017:1587:133-138. doi: 10.1007/978-1-4939-6892-3_13.

Abstract

Telomere dysfunctions, rendered through replicative attrition of telomeric DNA or due to the removal of shelterin components, are recognized as DNA double-stranded breaks (DSBs) by the DNA damage repair (DDR) pathway. This leads to the activation of DNA damage checkpoint sensors, including the Mre11-Rad50-Nbs1 (MRN) complex, γ-H2AX and 53BP1, the ATM and ATR signal-transducing kinases, and downstream effectors, including Chk1, Chk2, and p53. Robust DNA damage response signals at dysfunctional telomeres, achieved by the complete deletion of TRF2 or by expressing dominant-negative mutant TPP1ΔRD, can be detected by their association with γ-H2AX and 53BP1 forming "telomere dysfunction induced foci (TIFs)." Induction of TIFs at telomeres provides an opportunity to quantify the extent of telomere dysfunction and monitor downstream signaling pathways.

Keywords: DNA damage; Telomere dysfunction; Telomere-FISH; Telomere-induced foci.

MeSH terms

  • Animals
  • Ataxia Telangiectasia Mutated Proteins / genetics
  • Cell Cycle Proteins / genetics
  • Cells, Cultured
  • DNA / genetics
  • DNA Breaks, Double-Stranded
  • DNA Damage / genetics*
  • DNA Repair / genetics
  • DNA-Binding Proteins / genetics
  • HEK293 Cells
  • Humans
  • Mice
  • Signal Transduction / genetics
  • Telomere / genetics*
  • Tumor Suppressor p53-Binding Protein 1 / genetics

Substances

  • Cell Cycle Proteins
  • DNA-Binding Proteins
  • Tumor Suppressor p53-Binding Protein 1
  • DNA
  • Ataxia Telangiectasia Mutated Proteins