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      SAGA is a general cofactor for RNA polymerase II transcription

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          Summary

          Prior studies suggested that SAGA and TFIID are alternative factors that promote RNA polymerase II transcription with about 10% of genes in S. cerevisiae dependent on SAGA. We reassessed the role of SAGA by mapping its genome-wide location and role in global transcription in budding yeast. We find that SAGA maps to the UAS elements of most genes, overlapping with Mediator binding and irrespective of previous designations of SAGA or TFIID-dominated genes. Disruption of SAGA through mutation or rapid subunit depletion reduces transcription from nearly all genes, measured by newly-synthesized RNA. We also find that the acetyltransferase Gcn5 synergizes with Spt3 to promote global transcription and that Spt3 functions to stimulate TBP recruitment at all tested genes. Our data demonstrate that SAGA acts as a general cofactor required for essentially all RNA polymerase II transcription and is not consistent with the previous classification of SAGA and TFIID-dominated genes.

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          Most cited references53

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          Bidirectional promoters generate pervasive transcription in yeast.

          Genome-wide pervasive transcription has been reported in many eukaryotic organisms, revealing a highly interleaved transcriptome organization that involves hundreds of previously unknown non-coding RNAs. These recently identified transcripts either exist stably in cells (stable unannotated transcripts, SUTs) or are rapidly degraded by the RNA surveillance pathway (cryptic unstable transcripts, CUTs). One characteristic of pervasive transcription is the extensive overlap of SUTs and CUTs with previously annotated features, which prompts questions regarding how these transcripts are generated, and whether they exert function. Single-gene studies have shown that transcription of SUTs and CUTs can be functional, through mechanisms involving the generated RNAs or their generation itself. So far, a complete transcriptome architecture including SUTs and CUTs has not been described in any organism. Knowledge about the position and genome-wide arrangement of these transcripts will be instrumental in understanding their function. Here we provide a comprehensive analysis of these transcripts in the context of multiple conditions, a mutant of the exosome machinery and different strain backgrounds of Saccharomyces cerevisiae. We show that both SUTs and CUTs display distinct patterns of distribution at specific locations. Most of the newly identified transcripts initiate from nucleosome-free regions (NFRs) associated with the promoters of other transcripts (mostly protein-coding genes), or from NFRs at the 3' ends of protein-coding genes. Likewise, about half of all coding transcripts initiate from NFRs associated with promoters of other transcripts. These data change our view of how a genome is transcribed, indicating that bidirectionality is an inherent feature of promoters. Such an arrangement of divergent and overlapping transcripts may provide a mechanism for local spreading of regulatory signals-that is, coupling the transcriptional regulation of neighbouring genes by means of transcriptional interference or histone modification.
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            Dissecting the regulatory circuitry of a eukaryotic genome.

            Genome-wide expression analysis was used to identify genes whose expression depends on the functions of key components of the transcription initiation machinery in yeast. Components of the RNA polymerase II holoenzyme, the general transcription factor TFIID, and the SAGA chromatin modification complex were found to have roles in expression of distinct sets of genes. The results reveal an unanticipated level of regulation which is superimposed on that due to gene-specific transcription factors, a novel mechanism for coordinate regulation of specific sets of genes when cells encounter limiting nutrients, and evidence that the ultimate targets of signal transduction pathways can be identified within the initiation apparatus.
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              Nascent transcript sequencing visualizes transcription at nucleotide resolution.

              Recent studies of transcription have revealed a level of complexity not previously appreciated even a few years ago, both in the intricate use of post-initiation control and the mass production of rapidly degraded transcripts. Dissection of these pathways requires strategies for precisely following transcripts as they are being produced. Here we present an approach (native elongating transcript sequencing, NET-seq), based on deep sequencing of 3' ends of nascent transcripts associated with RNA polymerase, to monitor transcription at nucleotide resolution. Application of NET-seq in Saccharomyces cerevisiae reveals that although promoters are generally capable of divergent transcription, the Rpd3S deacetylation complex enforces strong directionality to most promoters by suppressing antisense transcript initiation. Our studies also reveal pervasive polymerase pausing and backtracking throughout the body of transcripts. Average pause density shows prominent peaks at each of the first four nucleosomes, with the peak location occurring in good agreement with in vitro biophysical measurements. Thus, nucleosome-induced pausing represents a major barrier to transcriptional elongation in vivo.
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                Author and article information

                Journal
                9802571
                20730
                Mol Cell
                Mol. Cell
                Molecular cell
                1097-2765
                1097-4164
                20 September 2017
                14 September 2017
                05 October 2017
                05 October 2018
                : 68
                : 1
                : 130-143.e5
                Affiliations
                [1 ]Institut de Génétique et de Biologie Moléculaire et Cellulaire, 67404 Illkirch, France
                [2 ]Centre National de la Recherche Scientifique, UMR7104, 67404 Illkirch, France
                [3 ]Institut National de la Santé et de la Recherche Médicale, U964, 67404 Illkirch, France
                [4 ]Université de Strasbourg, 67404 Illkirch, France
                [5 ]Basic Sciences Division, Fred Hutchinson Cancer Research Center, Seattle, WA, USA
                [6 ]Université Paris Diderot, Sorbonne Paris Cité, 75205 Paris, France
                [7 ]Molecular Cancer Research and Stem Cell Section, Regenerative Medicine Center and Center for Molecular Medicine, University Medical Center Utrecht c/o Hubrecht Institute, Uppsalalaan 8. 3584 CT Utrecht, The Netherlands
                [8 ]German Cancer Consortium (DKTK) partner site Freiburg, German Cancer Research Center (DKFZ) and Department of Urology, Medical Center-University of Freiburg, Germany
                Author notes
                [9]

                Lead Contact

                [* ]Correspondence: devys@ 123456igbmc.fr (D.D.), laszlo@ 123456igbmc.fr (L.T.)
                Article
                EMS74126
                10.1016/j.molcel.2017.08.016
                5632562
                28918903
                3e634359-f444-4eb0-ba91-5cdc93649233

                This manuscript version is made available under the CC-BY-NC-ND 4.0 license http://creativecommons.org/licenses/by-nc-nd/4.0/

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                Molecular biology
                Molecular biology

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