title: Purification of RNA Polymerase I-Associated Chromatin from Yeast Cells.
abstract: The native template of all eukaryotic nuclear RNA polymerases is chromatin. To understand how transcription occurs in vivo, it is important to define the chromatin environment of transcribing RNA Pols. Here, we describe a method used to characterize the distribution and the protein environment of RNA Pol I on ribosomal DNA during transcription in the yeast S. cerevisiae. The method is based on conventional chromatin immunoprecipitation and we propose quality control analyses at different steps of the procedure. Finally, the obtained samples are a useful source for downstream analyses by semiquantitative mass spectrometry or quantitative PCR.
authors: Universität Regensburg, Biochemie-Zentrum Regensburg (BZR), Lehrstuhl Biochemie I, 93053, Regensburg, Germany.
authors: Astrid
authors: A
authors: Bruckmann
authors: Universität Regensburg, Biochemie-Zentrum Regensburg (BZR), Lehrstuhl Biochemie I, 93053, Regensburg, Germany.
authors: Jan
authors: J
authors: Linnemann
authors: Universität Regensburg, Biochemie-Zentrum Regensburg (BZR), Lehrstuhl Biochemie I, 93053, Regensburg, Germany. jpfernandez@ur.de.
authors: Jorge
authors: J
authors: Perez-Fernandez
mesh_terms: D002843:Chromatin / Q000235:genetics / Q000302:isolation & purification* / Q000378:metabolism*; D047369:Chromatin Immunoprecipitation; D002846:Chromatography, Affinity / Q000379:methods; D004275:DNA, Ribosomal / Q000235:genetics; D059014:High-Throughput Nucleotide Sequencing; D020543:Proteome; D012318:RNA Polymerase I / Q000378:metabolism*; D012441:Saccharomyces cerevisiae / Q000235:genetics* / Q000378:metabolism*; D029701:Saccharomyces cerevisiae Proteins / Q000378:metabolism; D014158:Transcription, Genetic
chemical_list: D002843:Chromatin; D004275:DNA, Ribosomal; D020543:Proteome; D029701:Saccharomyces cerevisiae Proteins; D012318:RNA Polymerase I
keywords: Affinity purification; ChIP; Chromatin purification; Crosslink; Proteome-ChIP; RNA Polymerase I; Saccharomyces cerevisiae; Transcription; Yeast; rDNA; rRNA