Yeast TFIID serves as a coactivator for Rap1p by direct protein-protein interaction.
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.
Garbett
Krassimira A
KA
Department of Molecular Physiology and Biophysics, Vanderbilt University, School of Medicine, Nashville, TN 37232-0615, USA.
Tripathi
Manish K
MK
Cencki
Belgin
B
Layer
Justin H
JH
Weil
P Anthony
PA
D001667:Binding, Competitive; D004268:DNA-Binding Proteins / Q000737:chemistry; D004742:Enhancer Elements, Genetic; D015966:Gene Expression Regulation, Fungal; D011485:Protein Binding; D025941:Protein Interaction Mapping; D017434:Protein Structure, Tertiary; D011994:Recombinant Proteins / Q000737:chemistry; D012441:Saccharomyces cerevisiae / Q000235:genetics* / Q000378:metabolism; D029701:Saccharomyces cerevisiae Proteins / Q000737:chemistry / Q000378:metabolism / Q000502:physiology*; D000089804:Shelterin Complex; D035181:TATA-Box Binding Protein / Q000737:chemistry; D034501:Telomere-Binding Proteins / Q000737:chemistry / Q000378:metabolism / Q000502:physiology*; D035362:Transcription Factor TFIID / Q000737:chemistry* / Q000378:metabolism; D014157:Transcription Factors / Q000737:chemistry / Q000378:metabolism / Q000502:physiology*; D014158:Transcription, Genetic; D015533:Transcriptional Activation
D004268:DNA-Binding Proteins; C440829:RAP1 protein, S cerevisiae; D011994:Recombinant Proteins; D029701:Saccharomyces cerevisiae Proteins; D000089804:Shelterin Complex; D035181:TATA-Box Binding Protein; D034501:Telomere-Binding Proteins; D035362:Transcription Factor TFIID; D014157:Transcription Factors