title: Transcriptional analysis of purified histone acetyltransferase complexes.
abstract: Acetylation of lysine residues within the amino-terminal tails of the core histone proteins is strongly correlated to the regulation of gene transcription in vivo. To directly study the effects of histone acetylation on transcription, we have developed a biochemical system examining the regulation of RNA polymerase II-directed transcription by native histone acetyltransferases (HATs). For the promoter sequences investigated, it has been demonstrated that HATs facilitate transcription from nucleosomal DNA templates in an acetyl-CoA-dependent fashion but do not affect transcription from histone-free templates. Here, protocols are presented describing the in vitro assembly of evenly spaced nucleosomal arrays on DNA fragments harboring gene regulatory sequences and the use of these templates with purified HAT complexes in transcription assays.
authors.affiliation: Department of Biochemistry and Biophysics, University of California at San Francisco, 513 Parnassus Avenue, San Francisco, California 94143-0448, USA.
authors.forename: D J
authors.initials: DJ
authors.lastname: Steger
authors.forename: J L
authors.initials: JL
authors.lastname: Workman
mesh_terms: D000123:Acetyltransferases / Q000235:genetics* / Q000302:isolation & purification; D015971:Gene Expression Regulation, Enzymologic; D051548:Histone Acetyltransferases; D008967:Molecular Biology / Q000379:methods*; D029701:Saccharomyces cerevisiae Proteins; D014158:Transcription, Genetic
chemical_list: D029701:Saccharomyces cerevisiae Proteins; D000123:Acetyltransferases; D051548:Histone Acetyltransferases