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Molecular and Cellular Biology, January 2007, p. 297-311, Vol. 27, No. 1
0270-7306/07/$08.00+0     doi:10.1128/MCB.01558-06
Copyright © 2007, American Society for Microbiology. All Rights Reserved.

Yeast TFIID Serves as a Coactivator for Rap1p by Direct Protein-Protein Interaction{triangledown}

Krassimira A. Garbett, Manish K. Tripathi, Belgin Cencki, Justin H. Layer, and P. Anthony Weil*

Department of Molecular Physiology and Biophysics, Vanderbilt University, School of Medicine, Nashville, Tennessee 37232-0615

Received 21 August 2006/ Returned for modification 6 September 2006/ Accepted 13 October 2006

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 UASRAP1 enhancer-driven reporter gene requires both Rap1p and TFIID and is independent of the Fhl1p-Ifh1p coregulator. UASRAP1 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.

* Corresponding author. Mailing address: Department of Molecular Physiology and Biophysics, Vanderbilt University, School of Medicine, Nashville, TN 37232-0615. Phone: (615) 322-7007. Fax: (615) 322-7236. E-mail: tony.weil{at}vanderbilt.edu.

{triangledown} Published ahead of print on 30 October 2006.

Molecular and Cellular Biology, January 2007, p. 297-311, Vol. 27, No. 1
0270-7306/07/$08.00+0     doi:10.1128/MCB.01558-06
Copyright © 2007, American Society for Microbiology. All Rights Reserved.



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