title: Mediator, TATA-binding protein, and RNA polymerase II contribute to low histone occupancy at active gene promoters in yeast.
abstract: Transcription by RNA polymerase II (Pol II) in eukaryotes requires the Mediator complex, and often involves chromatin remodeling and histone eviction at active promoters. Here we address the role of Mediator in recruitment of the Swi/Snf chromatin remodeling complex and its role, along with components of the preinitiation complex (PIC), in histone eviction at inducible and constitutively active promoters in the budding yeast Saccharomyces cerevisiae. We show that recruitment of the Swi/Snf chromatin remodeling complex to the induced CHA1 promoter, as well as its association with several constitutively active promoters, depends on the Mediator complex but is independent of Mediator at the induced MET2 and MET6 genes. Although transcriptional activation and histone eviction at CHA1 depends on Swi/Snf, Swi/Snf recruitment is not sufficient for histone eviction at the induced CHA1 promoter. Loss of Swi/Snf activity does not affect histone occupancy of several constitutively active promoters; in contrast, higher histone occupancy is seen at these promoters in Mediator and PIC component mutants. We propose that an initial activator-dependent, nucleosome remodeling step allows PIC components to outcompete histones for occupancy of promoter sequences. We also observe reduced promoter association of Mediator and TATA-binding protein in a Pol II (rpb1-1) mutant, indicating mutually cooperative binding of these components of the transcription machinery and indicating that it is the PIC as a whole whose binding results in stable histone eviction.
authors.affiliation: From the Laboratory of Molecular Genetics, Wadsworth Center, New York State Department of Health, Albany, New York 12201-0509.
authors.forename: Suraiya A
authors.initials: SA
authors.lastname: Ansari
authors.affiliation: From the Laboratory of Molecular Genetics, Wadsworth Center, New York State Department of Health, Albany, New York 12201-0509.
authors.forename: Emily
authors.initials: E
authors.lastname: Paul
authors.affiliation: the Adolf-Butenandt-Institut, Universität München, 80336 Munich, Germany.
authors.forename: Sebastian
authors.initials: S
authors.lastname: Sommer
authors.affiliation: the Adolf-Butenandt-Institut, Universität München, 80336 Munich, Germany.
authors.forename: Corinna
authors.initials: C
authors.lastname: Lieleg
authors.affiliation: From the Laboratory of Molecular Genetics, Wadsworth Center, New York State Department of Health, Albany, New York 12201-0509, the Department of Biomedical Science, University at Albany School of Public Health, Albany, New York 12201-0509, and.
authors.forename: Qiye
authors.initials: Q
authors.lastname: He
authors.affiliation: From the Laboratory of Molecular Genetics, Wadsworth Center, New York State Department of Health, Albany, New York 12201-0509.
authors.forename: Alexandre Z
authors.initials: AZ
authors.lastname: Daly
authors.affiliation: From the Laboratory of Molecular Genetics, Wadsworth Center, New York State Department of Health, Albany, New York 12201-0509.
authors.forename: Kara A
authors.initials: KA
authors.lastname: Rode
authors.affiliation: From the Laboratory of Molecular Genetics, Wadsworth Center, New York State Department of Health, Albany, New York 12201-0509.
authors.forename: Wesley T
authors.initials: WT
authors.lastname: Barber
authors.affiliation: From the Laboratory of Molecular Genetics, Wadsworth Center, New York State Department of Health, Albany, New York 12201-0509.
authors.forename: Laura C
authors.initials: LC
authors.lastname: Ellis
authors.affiliation: From the Laboratory of Molecular Genetics, Wadsworth Center, New York State Department of Health, Albany, New York 12201-0509, the Department of Biomedical Science, University at Albany School of Public Health, Albany, New York 12201-0509, and.
authors.forename: Erika
authors.initials: E
authors.lastname: LaPorta
authors.affiliation: From the Laboratory of Molecular Genetics, Wadsworth Center, New York State Department of Health, Albany, New York 12201-0509.
authors.forename: Amanda M
authors.initials: AM
authors.lastname: Orzechowski
authors.affiliation: From the Laboratory of Molecular Genetics, Wadsworth Center, New York State Department of Health, Albany, New York 12201-0509.
authors.forename: Emily
authors.initials: E
authors.lastname: Taylor
authors.affiliation: From the Laboratory of Molecular Genetics, Wadsworth Center, New York State Department of Health, Albany, New York 12201-0509.
authors.forename: Tanner
authors.initials: T
authors.lastname: Reeb
authors.affiliation: From the Laboratory of Molecular Genetics, Wadsworth Center, New York State Department of Health, Albany, New York 12201-0509.
authors.forename: Jason
authors.initials: J
authors.lastname: Wong
authors.affiliation: the Adolf-Butenandt-Institut, Universität München, 80336 Munich, Germany.
authors.forename: Philipp
authors.initials: P
authors.lastname: Korber
authors.affiliation: From the Laboratory of Molecular Genetics, Wadsworth Center, New York State Department of Health, Albany, New York 12201-0509, the Department of Biomedical Science, University at Albany School of Public Health, Albany, New York 12201-0509, and randall.morse@wadsworth.org.
authors.forename: Randall H
authors.initials: RH
authors.lastname: Morse
mesh_terms: D050976:Basic-Leucine Zipper Transcription Factors / Q000235:genetics / Q000378:metabolism; D015152:Blotting, Northern; D002843:Chromatin / Q000235:genetics / Q000378:metabolism; D042002:Chromatin Assembly and Disassembly; D047369:Chromatin Immunoprecipitation; D002868:Chromosomal Proteins, Non-Histone / Q000235:genetics / Q000378:metabolism; D004268:DNA-Binding Proteins / Q000235:genetics / Q000378:metabolism; D006657:Histones / Q000378:metabolism*; D056892:Mediator Complex / Q000235:genetics / Q000378:metabolism*; D009154:Mutation; D009687:Nuclear Proteins / Q000235:genetics / Q000378:metabolism; D009707:Nucleosomes / Q000235:genetics / Q000378:metabolism; D011401:Promoter Regions, Genetic / Q000235:genetics*; D011485:Protein Binding; D012319:RNA Polymerase II / Q000235:genetics / Q000378:metabolism*; D012441:Saccharomyces cerevisiae / Q000235:genetics / Q000378:metabolism; D029701:Saccharomyces cerevisiae Proteins / Q000235:genetics / Q000378:metabolism*; D035181:TATA-Box Binding Protein / Q000235:genetics / Q000378:metabolism*; D014157:Transcription Factors / Q000235:genetics / Q000378:metabolism; D061785:Transcription Initiation, Genetic; D015533:Transcriptional Activation
chemical_list: D050976:Basic-Leucine Zipper Transcription Factors; D002843:Chromatin; D002868:Chromosomal Proteins, Non-Histone; D004268:DNA-Binding Proteins; D006657:Histones; C073675:MET4 protein, S cerevisiae; D056892:Mediator Complex; D009687:Nuclear Proteins; D009707:Nucleosomes; C066190:SNF5 protein, S cerevisiae; C072981:SWI3 protein, S cerevisiae; D029701:Saccharomyces cerevisiae Proteins; D035181:TATA-Box Binding Protein; D014157:Transcription Factors; D012319:RNA Polymerase II; C511736:RPB1 protein, S cerevisiae
keywords: Chromatin Immunoprecipitation (ChiP); Chromatin Remodeling; Gene Transcription; Mediator; Nucleosome; Saccharomyces Cerevisiae; Swi/Snf Complex; Transcription; Transcription Coactivators; Yeast Genetics
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