title: A functional module of yeast mediator that governs the dynamic range of heat-shock gene expression.
abstract: We report the results of a genetic screen designed to identify transcriptional coregulators of yeast heat-shock factor (HSF). This sequence-specific activator is required to stimulate both basal and induced transcription; however, the identity of factors that collaborate with HSF in governing noninduced heat-shock gene expression is unknown. In an effort to identify these factors, we isolated spontaneous extragenic suppressors of hsp82-deltaHSE1, an allele of HSP82 that bears a 32-bp deletion of its high-affinity HSF-binding site, yet retains its two low-affinity HSF sites. Nearly 200 suppressors of the null phenotype of hsp82-deltaHSE1 were isolated and characterized, and they sorted into six expression without heat-shock element (EWE) complementation groups. Strikingly, all six groups contain alleles of genes that encode subunits of Mediator. Three of the six subunits, Med7, Med10/Nut2, and Med21/Srb7, map to Mediator's middle domain; two subunits, Med14/Rgr1 and Med16/Sin4, to its tail domain; and one subunit, Med19/Rox3, to its head domain. Mutations in genes encoding these factors enhance not only the basal transcription of hsp82-deltaHSE1, but also that of wild-type heat-shock genes. In contrast to their effect on basal transcription, the more severe ewe mutations strongly reduce activated transcription, drastically diminishing the dynamic range of heat-shock gene expression. Notably, targeted deletion of other Mediator subunits, including the negative regulators Cdk8/Srb10, Med5/Nut1, and Med15/Gal11 fail to derepress hsp82-deltaHSE1. Taken together, our data suggest that the Ewe subunits constitute a distinct functional module within Mediator that modulates both basal and induced heat-shock gene transcription.
authors.affiliation: Department of Biochemistry and Molecular Biology, Louisiana State University Health Sciences Center, Shreveport, Louisiana 71130-3932, USA.
authors.forename: Harpreet
authors.initials: H
authors.lastname: Singh
authors.forename: Alexander M
authors.initials: AM
authors.lastname: Erkine
authors.forename: Selena B
authors.initials: SB
authors.lastname: Kremer
authors.forename: Harry M
authors.initials: HM
authors.lastname: Duttweiler
authors.forename: Donnie A
authors.initials: DA
authors.lastname: Davis
authors.forename: Jabed
authors.initials: J
authors.lastname: Iqbal
authors.forename: Rachel R
authors.initials: RR
authors.lastname: Gross
authors.forename: David S
authors.initials: DS
authors.lastname: Gross
mesh_terms: D000483:Alleles; D017353:Gene Deletion; D015966:Gene Expression Regulation, Fungal; D005800:Genes, Fungal; D005816:Genetic Complementation Test; D008040:Genetic Linkage; D006360:Heat-Shock Proteins / Q000378:metabolism*; D056892:Mediator Complex; D009154:Mutation; D010641:Phenotype; D029701:Saccharomyces cerevisiae Proteins / Q000235:genetics*; D015534:Trans-Activators / Q000235:genetics*; D014158:Transcription, Genetic; D001616:beta-Galactosidase / Q000378:metabolism
chemical_list: D006360:Heat-Shock Proteins; C110184:MED7 protein, S cerevisiae; D056892:Mediator Complex; D029701:Saccharomyces cerevisiae Proteins; D015534:Trans-Activators; D001616:beta-Galactosidase