title: Identification of telomere-associated molecules by engineered DNA-binding molecule-mediated chromatin immunoprecipitation (enChIP).
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.
authors.affiliation: Combined Program on Microbiology and Immunology, Research Institute for Microbial Diseases, Osaka University, 3-1 Yamadaoka, Suita, 565-0871 Osaka, Japan.
authors.forename: Toshitsugu
authors.initials: T
authors.lastname: Fujita
authors.forename: Yoshinori
authors.initials: Y
authors.lastname: Asano
authors.forename: Junko
authors.initials: J
authors.lastname: Ohtsuka
authors.forename: Yoko
authors.initials: Y
authors.lastname: Takada
authors.forename: Kazunobu
authors.initials: K
authors.lastname: Saito
authors.forename: Rieko
authors.initials: R
authors.lastname: Ohki
authors.forename: Hodaka
authors.initials: H
authors.lastname: Fujii
mesh_terms: D001483:Base Sequence; D001665:Binding Sites; D002460:Cell Line; D047369:Chromatin Immunoprecipitation / Q000379:methods*; D004268:DNA-Binding Proteins / Q000378:metabolism*; D006801:Humans; D059372:Nucleotide Motifs; D011485:Protein Binding; D021381:Protein Transport; D012313:RNA / Q000235:genetics / Q000378:metabolism; D019098:Telomerase / Q000235:genetics; D016615:Telomere / Q000235:genetics / Q000378:metabolism*; D034501:Telomere-Binding Proteins / Q000378:metabolism*
chemical_list: D004268:DNA-Binding Proteins; D034501:Telomere-Binding Proteins; D012313:RNA; D019098:Telomerase