Identification of telomere-associated molecules by engineered DNA-binding molecule-mediated chromatin immunoprecipitation (enChIP)

Sci Rep. 2013 Nov 8:3:3171. doi: 10.1038/srep03171.

Abstract

Biochemical analysis of molecular interactions in specific genomic regions requires their isolation while retaining molecular interactions in vivo. Here, we report isolation of telomeres by engineered DNA-binding molecule-mediated chromatin immunoprecipitation (enChIP) using a transcription activator-like (TAL) protein recognizing telomere repeats. Telomeres recognized by the tagged TAL protein were immunoprecipitated with an antibody against the tag and subjected to identification of telomere-binding molecules. enChIP-mass spectrometry (enChIP-MS) targeting telomeres identified known and novel telomere-binding proteins. The data have been deposited to the ProteomeXchange with identifier PXD000461. In addition, we showed that RNA associated with telomeres could be isolated by enChIP. Identified telomere-binding molecules may play important roles in telomere biology. enChIP using TAL proteins would be a useful tool for biochemical analysis of specific genomic regions of interest.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Base Sequence
  • Binding Sites
  • Cell Line
  • Chromatin Immunoprecipitation / methods*
  • DNA-Binding Proteins / metabolism*
  • Humans
  • Nucleotide Motifs
  • Protein Binding
  • Protein Transport
  • RNA / genetics
  • RNA / metabolism
  • Telomerase / genetics
  • Telomere / genetics
  • Telomere / metabolism*
  • Telomere-Binding Proteins / metabolism*

Substances

  • DNA-Binding Proteins
  • Telomere-Binding Proteins
  • RNA
  • Telomerase