title: Facilitated assembly of the preinitiation complex by separated tail and head/middle modules of the mediator.
abstract: Mediator is a general coactivator of RNA polymerase II (RNA pol II) bridging enhancer-bound transcriptional factors with RNA pol II. Mediator is organized in three distinct subcomplexes: head, middle, and tail modules. The head and middle modules interact with RNA pol II, and the tail module interacts with transcriptional activators. Deletion of one of the tail subunits SIN4 results in derepression of a subset of genes, including FLR1, by a largely unknown mechanism. Here we show that derepression of FLR1 transcription in sin4Δ cells occurs by enhanced recruitment of the mediator as well as Swi/Snf and SAGA complexes. The tail and head/middle modules of the mediator behave as separate complexes at the induced FLR1 promoter. While the tail module remains anchored to the promoter, the head/middle modules are also found in the coding region. The separation of the tail and head/middle modules in sin4Δ cells is also supported by the altered stoichiometry of the tail and head/middle modules at several tested promoters. Deletion of another subunit of the tail module MED2 in sin4Δ cells results in significantly decreased transcription of FLR1, pointing to the importance of the integrity of the separated tail module in derepression. All tested genes exhibited increased recruitment of the tail domain; however, only genes with increased occupancy of the head/middle modules also displayed increased transcription. The separated tail module thus represents a promiscuous transcriptional factor that binds to many different promoters and is necessary for derepression of FLR1 in sin4Δ cells.
authors.affiliation: Department of Biological Sciences, St. John's University, 8000 Utopia Parkway, Queens, NY 11439, USA.
authors.forename: Luciano
authors.initials: L
authors.lastname: Galdieri
authors.forename: Parima
authors.initials: P
authors.lastname: Desai
authors.forename: Ales
authors.initials: A
authors.lastname: Vancura
mesh_terms: D004271:DNA, Fungal / Q000378:metabolism; D017353:Gene Deletion; D056892:Mediator Complex / Q000235:genetics / Q000378:metabolism*; D008954:Models, Biological; D027361:Organic Anion Transporters / Q000096:biosynthesis*; D011401:Promoter Regions, Genetic; D011485:Protein Binding; D021122:Protein Subunits / Q000235:genetics / Q000378:metabolism; D012441:Saccharomyces cerevisiae / Q000201:enzymology*; D029701:Saccharomyces cerevisiae Proteins / Q000096:biosynthesis* / Q000235:genetics*; D014158:Transcription, Genetic
chemical_list: D004271:DNA, Fungal; C118429:FLR1 protein, S cerevisiae; D056892:Mediator Complex; D027361:Organic Anion Transporters; D021122:Protein Subunits; C076878:SIN4 protein, S cerevisiae; D029701:Saccharomyces cerevisiae Proteins