title: Direct identification of insulator components by insertional chromatin immunoprecipitation.
abstract: Comprehensive understanding of mechanisms of epigenetic regulation requires identification of molecules bound to genomic regions of interest in vivo. However, non-biased methods to identify molecules bound to specific genomic loci in vivo are limited. Here, we applied insertional chromatin immunoprecipitation (iChIP) to direct identification of components of insulator complexes, which function as boundaries of chromatin domain. We found that the chicken β-globin HS4 (cHS4) insulator complex contains an RNA helicase protein, p68/DDX5; an RNA species, steroid receptor RNA activator 1; and a nuclear matrix protein, Matrin-3, in vivo. Binding of p68 and Matrin-3 to the cHS4 insulator core sequence was mediated by CCCTC-binding factor (CTCF). Thus, our results showed that it is feasible to directly identify proteins and RNA bound to a specific genomic region in vivo by using iChIP.
authors: Combined Program on Microbiology and Immunology, Research Institute for Microbial Diseases, Osaka University, Osaka, Japan.
authors: Toshitsugu
authors: T
authors: Fujita
authors: Hodaka
authors: H
authors: Fujii
mesh_terms: D000818:Animals; D001665:Binding Sites; D002645:Chickens; D047369:Chromatin Immunoprecipitation / Q000379:methods*; D005786:Gene Expression Regulation; D016678:Genome; D016254:Mutagenesis, Insertional; D011506:Proteins / Q000032:analysis*; D012313:RNA / Q000032:analysis*
chemical_list: D011506:Proteins; D012313:RNA