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Molecular and Cellular Biology, October 2001, p. 6668-6680, Vol. 21, No. 19
0270-7306/01/$04.00+0   DOI: 10.1128/MCB.21.19.6668-6680.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.

Molecular Genetic Dissection of TAF25, an Essential Yeast Gene Encoding a Subunit Shared by TFIID and SAGA Multiprotein Transcription Factors

Jay Kirchner, Steven L. Sanders, Edward Klebanow,dagger and P. Anthony Weil*

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

Received 30 April 2001/Returned for modification 31 May 2001/Accepted 27 June 2001

We have performed a systematic structure-function analysis of Saccharomyces cerevisiae TAF25, an evolutionarily conserved, single-copy essential gene which encodes the 206-amino-acid TAF25p protein. TAF25p is an integral subunit of both the 15-subunit general transcription factor TFIID and the multisubunit, chromatin-acetylating transcriptional coactivator SAGA. We used hydroxylamine mutagenesis, targeted deletion, alanine-scanning mutagenesis, high-copy suppression methods, and two-hybrid screening to dissect TAF25. Temperature-sensitive mutant strains generated were used for coimmunoprecipitation and transcription analyses to define the in vivo functions of TAF25p. The results of these analyses show that TAF25p is comprised of multiple mutable elements which contribute importantly to RNA polymerase II-mediated mRNA 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}mcmail.vanderbilt.edu.

dagger Present address: Argus Research Corporation, New York, NY 10006.

Molecular and Cellular Biology, October 2001, p. 6668-6680, Vol. 21, No. 19
0270-7306/01/$04.00+0   DOI: 10.1128/MCB.21.19.6668-6680.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.


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