Abstract
In vivo studies have previously shown that Saccharomyces cerevisiae ribosomal protein (RP) gene expression is controlled by the transcription factor repressor activator protein 1 (Rap1p) in a TFIID-dependent fashion. Here we have tested the hypothesis that yeast TFIID serves as a coactivator for RP gene transcription by directly interacting with Rap1p. We have found that purified recombinant Rap1p specifically interacts with purified TFIID in pull-down assays, and we have mapped the domains of Rap1p and subunits of TFIID responsible. In vitro transcription of a UAS(RAP1) enhancer-driven reporter gene requires both Rap1p and TFIID and is independent of the Fhl1p-Ifh1p coregulator. UAS(RAP1) enhancer-driven transactivation in extracts depleted of both Rap1p and TFIID is efficiently rescued by addition of physiological amounts of these two purified factors but not TATA-binding protein. We conclude that Rap1p and TFIID directly interact and that this interaction contributes importantly to RP gene transcription.
Publication types
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Research Support, N.I.H., Extramural
MeSH terms
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Binding, Competitive
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DNA-Binding Proteins / chemistry
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Enhancer Elements, Genetic
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Gene Expression Regulation, Fungal*
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Protein Binding
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Protein Interaction Mapping
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Protein Structure, Tertiary
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Recombinant Proteins / chemistry
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Saccharomyces cerevisiae / genetics*
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Saccharomyces cerevisiae / metabolism
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Saccharomyces cerevisiae Proteins / chemistry
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Saccharomyces cerevisiae Proteins / metabolism
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Saccharomyces cerevisiae Proteins / physiology*
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Shelterin Complex
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TATA-Box Binding Protein / chemistry
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Telomere-Binding Proteins / chemistry
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Telomere-Binding Proteins / metabolism
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Telomere-Binding Proteins / physiology*
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Transcription Factor TFIID / chemistry*
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Transcription Factor TFIID / metabolism
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Transcription Factors / chemistry
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Transcription Factors / metabolism
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Transcription Factors / physiology*
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Transcription, Genetic
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Transcriptional Activation
Substances
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DNA-Binding Proteins
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RAP1 protein, S cerevisiae
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Recombinant Proteins
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Saccharomyces cerevisiae Proteins
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Shelterin Complex
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TATA-Box Binding Protein
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Telomere-Binding Proteins
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Transcription Factor TFIID
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Transcription Factors