title: Live-cell studies of p300/CBP histone acetyltransferase activity and inhibition.
abstract: Histone acetyltransferase enzymes (HATs) are important therapeutic targets, but there are few cell-based assays available for evaluating the pharmacodynamics of HAT inhibitors. Here we present the application of a FRET-based reporter, Histac, in live-cell studies of p300/CBP HAT inhibition, by both genetic and pharmacologic disruption. shRNA knockdown of p300/CBP led to increased Histac FRET, thus suggesting a role for p300/CBP in the acetylation of the histone H4 tail. Additionally, we describe a new p300/CBP HAT inhibitor, C107, and show that it can also increase cellular Histac FRET. Taken together, these studies provide a live-cell strategy for identifying and evaluating p300/CBP inhibitors.
authors.affiliation: Department of Pharmacology and Molecular Sciences, The Johns Hopkins University School of Medicine, 725 North Wolfe Street, Baltimore, MD 21205, USA.
authors.forename: Beverley M
authors.initials: BM
authors.lastname: Dancy
authors.forename: Nicholas T
authors.initials: NT
authors.lastname: Crump
authors.forename: Daniel J
authors.initials: DJ
authors.lastname: Peterson
authors.forename: Chandrani
authors.initials: C
authors.lastname: Mukherjee
authors.forename: Erin M
authors.initials: EM
authors.lastname: Bowers
authors.forename: Young-Hoon
authors.initials: YH
authors.lastname: Ahn
authors.forename: Minoru
authors.initials: M
authors.lastname: Yoshida
authors.forename: Jin
authors.initials: J
authors.lastname: Zhang
authors.forename: Louis C
authors.initials: LC
authors.lastname: Mahadevan
authors.forename: David J
authors.initials: DJ
authors.lastname: Meyers
authors.forename: Jef D
authors.initials: JD
authors.lastname: Boeke
authors.forename: Philip A
authors.initials: PA
authors.lastname: Cole
mesh_terms: D000107:Acetylation; D000818:Animals; D019556:COS Cells; D002470:Cell Survival / Q000187:drug effects; D002522:Chlorocebus aethiops; D004791:Enzyme Inhibitors / Q000138:chemical synthesis / Q000737:chemistry / Q000494:pharmacology; D031541:Fluorescence Resonance Energy Transfer; D034622:RNA Interference; D034741:RNA, Small Interfering / Q000378:metabolism; D011994:Recombinant Proteins / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism; D050880:p300-CBP Transcription Factors / Q000037:antagonists & inhibitors* / Q000235:genetics / Q000378:metabolism; D000099224:p300-CBP-Associated Factor
chemical_list: D004791:Enzyme Inhibitors; D034741:RNA, Small Interfering; D011994:Recombinant Proteins; D050880:p300-CBP Transcription Factors; D000099224:p300-CBP-Associated Factor