Direct Transactivator-Transcription Factor IID (TFIID) Contacts Drive Yeast Ribosomal Protein Gene Transcription

• Published in: The Journal of biological chemistry 2010-05, Vol.285 (20), p.15489-15499
• Main Authors: Layer, Justin H., Miller, Scott G., Weil, P. Anthony
• Format: Article
• Language: English
• Subjects: Binding Sites; Coactivator; Coregulator Transcription; Fungal Proteins - genetics; Gene Regulation; Gene Transcription; General Transcription Factors; Polymerase Chain Reaction; Protein Binding; Rap1; Ribosomal Proteins - genetics; TBP-associated Factor; TFIID; Trans-Activators - metabolism; Transcription Coactivators; Transcription Factor TFIID - metabolism; Transcription Factors; Transcription Regulation; Transcription, Genetic
• ISSN: 0021-9258; 1083-351X
• DOI: 10.1074/jbc.M110.104810

Transcription factor IID (TFIID) plays a key role in regulating eukaryotic gene expression by directly binding promoters and enhancer-bound transactivator proteins. However, the precise mechanisms and outcomes of transactivator-TFIID interaction remain unclear. Transcription of yeast ribosomal protein genes requires TFIID and the DNA-binding transactivator Rap1. We have previously shown that Rap1 directly binds to the TFIID complex through interaction with its TATA-binding protein-associated factor (Taf) subunits Taf4, -5, and -12. Here, we identify and characterize the Rap1 binding domains (RBDs) of Taf4 and Taf5. These RBDs are essential for viability but dispensable for Taf-Taf interactions and TFIID stability. Cells expressing altered Rap1 binding domains exhibit conditional growth, synthetic phenotypes when expressed in combination or with altered Rap1, and are selectively defective in ribosomal protein gene transcription. Taf4 and Taf5 proteins with altered RBDs bind Rap1 with reduced affinity. We propose that collectively the Taf4, Taf5, and Taf12 subunits of TFIID represent the physical and functional targets for Rap1 interaction and, furthermore, that these interactions drive ribosomal protein gene transcription.