title: Assessing HDAC Function in the Regulation of Signal Transducer and Activator of Transcription 5 (STAT5) Activity Using Chromatin Immunoprecipitation (ChIP).
abstract: Transcriptional activation by STAT5 is repressed by deacetylase inhibitors. Investigating the role of deacetylases (HDACs) in STAT5-mediated transcription implies the analysis of molecular events taking place at the chromatin level. We describe here two alternative methods of chromatin immunoprecipitation that allow the characterization of chromatin modifications ensuing STAT5 activation and its inhibition by deacetylase inhibitors, in particular changes in histone acetylation, in histone occupancy, and in the association/dissociation of transcription factors and other chromatin-associated factors.
authors.affiliation: Stat5 Signaling Research Group, Institute of Immunology, University of Regensburg, Franz-Josef-Strauss-Allee 11, 93053, Regensburg, Germany.
authors.forename: Sophia
authors.initials: S
authors.lastname: Pinz
authors.affiliation: Stat5 Signaling Research Group, Institute of Immunology, University of Regensburg, Franz-Josef-Strauss-Allee 11, 93053, Regensburg, Germany. anne.rascle@klinik.uni-regensburg.de.
authors.forename: Anne
authors.initials: A
authors.lastname: Rascle
mesh_terms: D000107:Acetylation; D000818:Animals; D001402:B-Lymphocytes / Q000166:cytology / Q000187:drug effects / Q000276:immunology*; D045744:Cell Line, Tumor; D002843:Chromatin / Q000737:chemistry / Q000276:immunology; D047369:Chromatin Immunoprecipitation; D002868:Chromosomal Proteins, Non-Histone / Q000235:genetics / Q000276:immunology; D044127:Epigenesis, Genetic; D056572:Histone Deacetylase Inhibitors / Q000494:pharmacology*; D006655:Histone Deacetylases / Q000235:genetics* / Q000276:immunology; D006657:Histones / Q000235:genetics / Q000276:immunology; D006877:Hydroxamic Acids / Q000494:pharmacology*; D007377:Interleukin-3 / Q000494:pharmacology; D008213:Lymphocyte Activation; D051379:Mice; D050799:STAT5 Transcription Factor / Q000235:genetics* / Q000276:immunology; D014157:Transcription Factors; D014158:Transcription, Genetic; D000092468:Bromodomain Containing Proteins
chemical_list: D002843:Chromatin; D002868:Chromosomal Proteins, Non-Histone; D056572:Histone Deacetylase Inhibitors; D006655:Histone Deacetylases; D006657:Histones; D006877:Hydroxamic Acids; D007377:Interleukin-3; D050799:STAT5 Transcription Factor; D014157:Transcription Factors; C490844:Brd2 protein, mouse; D000092468:Bromodomain Containing Proteins; C012589:trichostatin A
keywords: Brd2; ChIP; Chromatin immunoprecipitation; Deacetylase; Deacetylase inhibitor; HDAC; Histone acetylation; MNase digestion; Micrococcal nuclease; STAT5; Sonication; TSA; Trichostatin A