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Molecular and Cellular Biology, October 2001, p. 6668-6680, Vol. 21, No. 19
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.
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
and
*
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.
Present address: Argus Research Corporation, New York, NY 10006.
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