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      Mediator protein mutations that selectively abolish activated transcription.

      Proceedings of the National Academy of Sciences of the United States of America
      Cell-Free System, DNA-Binding Proteins, Fungal Proteins, metabolism, Gene Deletion, Gene Expression Regulation, Fungal, Models, Genetic, Mutation, Protein Kinases, Saccharomyces cerevisiae Proteins, Structure-Activity Relationship, Trans-Activators, genetics, Transcription, Genetic, Transcriptional Activation, Yeasts

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          Abstract

          Deletion of any one of three subunits of the yeast Mediator of transcriptional regulation, Med2, Pgd1 (Hrs1), and Sin4, abolished activation by Gal4-VP16 in vitro. By contrast, other Mediator functions, stimulation of basal transcription and of TFIIH kinase activity, were unaffected. A different but overlapping Mediator subunit dependence was found for activation by Gcn4. The genetic requirements for activation in vivo were closely coincident with those in vitro. A whole genome expression profile of a Deltamed2 strain showed diminished transcription of a subset of inducible genes but only minor effects on "basal" transcription. These findings make an important connection between transcriptional activation in vitro and in vivo, and identify Mediator as a "global" transcriptional coactivator.

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