title: Similar temporal and spatial recruitment of native 19S and 20S proteasome subunits to transcriptionally active chromatin.
abstract: It has recently become clear that components of the proteasome are recruited to sites of gene transcription. Prevailing evidence suggests that the transcriptionally relevant form of the proteasome is a subcomplex of 19S base proteins, which functions as an ATP-dependent chaperone that influences transcriptional processes. Despite this notion, compelling evidence for a transcription-dedicated 19S base complex is lacking, and 20S proteasome subunits have been shown to associate with chromatin in some contexts. To gain insight into the form of the proteasome that is recruited to chromatin, we assembled a panel of highly specific antibodies that recognize native yeast proteasome subunits in chromatin immunoprecipitation assays. Using these reagents, we show that components from the three major subassemblies of the proteasome--19S lid, 19S base, and 20S core--associate with the activated GAL10 gene in yeast in a virtually indistinguishable manner. We find that proteasome subunits Rpt1, Rpt4, Rpn8, Rpn12, Pre6, and Pre10 are recruited to GAL10 rapidly upon galactose induction. These subunits associate with the entire transcribed portion of GAL10, display near-identical patterns of distribution, and dissociate from chromatin rapidly once transcription is shut down. We also find that proteasome subunits are enriched at telomeres and at genes transcribed by RNA polymerase III. Our data suggest that the transcriptionally relevant form of the proteasome is the canonical 26S complex.
authors.affiliation: Department of Cell and Developmental Biology, Vanderbilt University School of Medicine, 465 21st Avenue South, Nashville, TN 37232, USA.
authors.forename: Fuqiang
authors.initials: F
authors.lastname: Geng
authors.forename: William P
authors.initials: WP
authors.lastname: Tansey
mesh_terms: D000251:Adenosine Triphosphatases / Q000235:genetics / Q000378:metabolism; D015153:Blotting, Western; D002843:Chromatin / Q000235:genetics / Q000378:metabolism*; D047369:Chromatin Immunoprecipitation; D005690:Galactose / Q000494:pharmacology; D009154:Mutation; D046988:Proteasome Endopeptidase Complex / Q000235:genetics / Q000378:metabolism*; D011485:Protein Binding; D021122:Protein Subunits / Q000235:genetics / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012441:Saccharomyces cerevisiae / Q000235:genetics / Q000378:metabolism*; D029701:Saccharomyces cerevisiae Proteins / Q000235:genetics / Q000378:metabolism*; D013997:Time Factors; D015534:Trans-Activators / Q000235:genetics / Q000378:metabolism; D014158:Transcription, Genetic / Q000187:drug effects
chemical_list: D002843:Chromatin; C518355:GAL10 protein, S cerevisiae; D021122:Protein Subunits; C074096:RPN12 protein, S cerevisiae; D029701:Saccharomyces cerevisiae Proteins; D015534:Trans-Activators; C573323:Pre10 protein, S cerevisiae; C573322:Pre6 protein, S cerevisiae; D046988:Proteasome Endopeptidase Complex; C573321:Rpn8 protein, S cerevisiae; D000251:Adenosine Triphosphatases; C103015:RPT4 protein, S cerevisiae; C084493:RPT1 protein, S cerevisiae; D005690:Galactose