title: Real-time imaging of histone H4K12-specific acetylation determines the modes of action of histone deacetylase and bromodomain inhibitors.
abstract: Histone acetylation constitutes an epigenetic mark for transcriptional regulation. Here we developed a fluorescent probe to visualize acetylation of histone H4 Lys12 (H4K12) in living cells using fluorescence resonance energy transfer (FRET) and the binding of the BRD2 bromodomain to acetylated H4K12. Using this probe designated as Histac-K12, we demonstrated that histone H4K12 acetylation is retained in mitosis and that some histone deacetylase (HDAC) inhibitors continue to inhibit cellular HDAC activity even after their removal from the culture. In addition, a small molecule that interferes with ability of the bromodomain to bind to acetylated H4K12 could be assessed using Histac-K12 in cells. Thus, Histac-K12 will serve as a powerful tool not only to understand the dynamics of H4K12-specific acetylation but also to characterize small molecules that modulate the acetylation or interaction status of histones.
authors.affiliation: Chemical Genetics Laboratory, RIKEN Advanced Science Institute, Wako, Saitama 351-0198, Japan.
authors.forename: Tamaki
authors.initials: T
authors.lastname: Ito
authors.forename: Takashi
authors.initials: T
authors.lastname: Umehara
authors.forename: Kazuki
authors.initials: K
authors.lastname: Sasaki
authors.forename: Yoshihiro
authors.initials: Y
authors.lastname: Nakamura
authors.forename: Norikazu
authors.initials: N
authors.lastname: Nishino
authors.forename: Takaho
authors.initials: T
authors.lastname: Terada
authors.forename: Mikako
authors.initials: M
authors.lastname: Shirouzu
authors.forename: Balasundaram
authors.initials: B
authors.lastname: Padmanabhan
authors.forename: Shigeyuki
authors.initials: S
authors.lastname: Yokoyama
authors.forename: Akihiro
authors.initials: A
authors.lastname: Ito
authors.forename: Minoru
authors.initials: M
authors.lastname: Yoshida
mesh_terms: D000107:Acetylation / Q000187:drug effects; D000818:Animals; D001665:Binding Sites; D019556:COS Cells; D002470:Cell Survival; D002522:Chlorocebus aethiops; D002875:Chromosomes / Q000187:drug effects / Q000378:metabolism; D015195:Drug Design; D005456:Fluorescent Dyes / Q000378:metabolism; D056572:Histone Deacetylase Inhibitors / Q000494:pharmacology*; D006655:Histone Deacetylases / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D006657:Histones / Q000737:chemistry* / Q000378:metabolism*; D007700:Kinetics; D008239:Lysine; D008938:Mitosis / Q000187:drug effects; D008958:Models, Molecular; D057054:Molecular Imaging; D009154:Mutation; D017434:Protein Structure, Tertiary / Q000187:drug effects; D013379:Substrate Specificity; D013997:Time Factors; D014158:Transcription, Genetic / Q000187:drug effects
chemical_list: D005456:Fluorescent Dyes; D056572:Histone Deacetylase Inhibitors; D006657:Histones; D006655:Histone Deacetylases; D008239:Lysine