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Molecular and Cellular Biology, October 1998, p. 5861-5867, Vol. 18, No. 10
Department of Biochemistry and Molecular
Biology, University of New Hampshire, Durham, New Hampshire
03824,1 and
Department of Molecular
Physiology and Biophysics, Vanderbilt University School of
Medicine, Nashville, Tennessee 37232-06152
Received 22 April 1998/Returned for modification 15 June
1998/Accepted 25 June 1998
The yeast transcriptional activator ADR1, which is required for
ADH2 and other genes' expression, contains four
transactivation domains (TADs). While previous studies have shown that
these TADs act through GCN5 and ADA2, and presumably TFIIB, other
factors are likely to be involved in ADR1 function. In this study, we addressed the question of whether TFIID is also required for ADR1 action. In vitro binding studies indicated that TADI of ADR1 was able
to retain TAFII90 from yeast extracts and TADII could
retain TBP and TAFII130/145. TADIV, however, was capable of
retaining multiple TAFIIs, suggesting that TADIV was
binding TFIID from yeast whole-cell extracts. The ability of TADIV
truncation derivatives to interact with TFIID correlated with their
transcription activation potential in vivo. In addition, the ability of
LexA-ADR1-TADIV to activate transcription in vivo was compromised by a
mutation in TAFII130/145. ADR1 was found to associate in
vivo with TFIID in that immunoprecipitation of either
TAFII90 or TBP from yeast whole-cell extracts specifically
coimmunoprecipitated ADR1. Most importantly, depletion of
TAFII90 from yeast cells dramatically reduced
ADH2 derepression. These results indicate that ADR1
physically associates with TFIID and that its ability to activate
transcription requires an intact TFIID complex.
0270-7306/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.
ADR1-Mediated Transcriptional Activation Requires
the Presence of an Intact TFIID Complex
*
Corresponding author. Mailing address: Department of
Biochemistry and Molecular Biology, Rudman Hall, University of New
Hampshire, Durham, NH 03824. Phone: (603) 862-2427. Fax: (603)
862-4013. E-mail: cldenis{at}christa.unh.edu.
Scientific contribution 1993 from the New Hampshire Agricultural
Experiment Station.
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