title: PUB-NChIP--"in vivo biotinylation" approach to study chromatin in proximity to a protein of interest.
abstract: We have developed an approach termed PUB-NChIP (proximity utilizing biotinylation with native ChIP) to purify and study the protein composition of chromatin in proximity to a nuclear protein of interest. It is based on coexpression of (1) a protein of interest, fused with the bacterial biotin ligase BirA, together with (2) a histone fused to a biotin acceptor peptide (BAP), which is specifically biotinylated by BirA-fusion in the proximity of the protein of interest. Using the RAD18 protein as a model, we demonstrate that the RAD18-proximal chromatin is enriched in some H4 acetylated species. Moreover, the RAD18-proximal chromatin containing a replacement histone H2AZ has a different pattern of H4 acetylation. Finally, biotin pulse-chase experiments show that the H4 acetylation pattern starts to resemble the acetylation pattern of total H4 after the proximity of chromatin to RAD18 has been lost.
authors.affiliation: UMR8126, Université Paris-Sud 11, CNRS, Institut de Cancérologie Gustave Roussy, 94805 Villejuif, France.
authors.forename: Muhammad
authors.initials: M
authors.lastname: Shoaib
authors.forename: Arman
authors.initials: A
authors.lastname: Kulyyassov
authors.forename: Chloé
authors.initials: C
authors.lastname: Robin
authors.forename: Kinga
authors.initials: K
authors.lastname: Winczura
authors.forename: Pavel
authors.initials: P
authors.lastname: Tarlykov
authors.forename: Emmanuelle
authors.initials: E
authors.lastname: Despas
authors.forename: Patricia
authors.initials: P
authors.lastname: Kannouche
authors.forename: Erlan
authors.initials: E
authors.lastname: Ramanculov
authors.forename: Marc
authors.initials: M
authors.lastname: Lipinski
authors.forename: Vasily
authors.initials: V
authors.lastname: Ogryzko
mesh_terms: D000107:Acetylation; D019921:Biotinylation / Q000379:methods; D002460:Cell Line; D002843:Chromatin / Q000378:metabolism*; D047369:Chromatin Immunoprecipitation / Q000379:methods*; D006657:Histones; D006801:Humans; D009687:Nuclear Proteins / Q000378:metabolism*; D011499:Protein Processing, Post-Translational
chemical_list: D002843:Chromatin; D006657:Histones; D009687:Nuclear Proteins