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.

Keywords: Affinity purification; ChIP; Chromatin purification; Crosslink; Proteome-ChIP; RNA Polymerase I; Saccharomyces cerevisiae; Transcription; Yeast; rDNA; rRNA.