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Originally published In Press as doi:10.1074/jbc.M102416200 on May 3, 2001

J. Biol. Chem., Vol. 276, Issue 28, 25715-25726, July 13, 2001
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Requirement for Yeast TAF145 Function in Transcriptional Activation of the RPS5 Promoter That Depends on Both Core Promoter Structure and Upstream Activating Sequences*

Yoshihiro Tsukihashi, Masashi Kawaichi, and Tetsuro KokuboDagger

From the Division of Gene Function in Animals, Nara Institute of Science and Technology, 8916-5 Takayama, Ikoma, Nara 630-0101, Japan

The general transcription factor TFIID has been shown to be involved in both core promoter recognition and the transcriptional activation of eukaryotic genes. We recently isolated TAF145 (one of TFIID subunits) temperature-sensitive mutants in yeast, in which transcription of the TUB2 gene is impaired at restrictive temperatures due to a defect in core promoter recognition. Here, we show in these mutants that the transcription of the RPS5 gene is impaired, mostly due to a defect in transcriptional activation rather than to a defect in core promoter recognition, although the latter is slightly affected as well. Surprisingly, the RPS5 core promoter can be activated by various activation domains fused to a GAL4 DNA binding domain, but not by the original upstream activating sequence (UAS) of the RPS5 gene. In addition, a heterologous CYC1 core promoter can be activated by RPS5-UAS at normal levels even in these mutants. These observations indicate that a distinct combination of core promoters and activators may exploit alternative activation pathways that vary in their requirement for TAF145 function. In addition, a particular function of TAF145 that is deleted in our mutants appears to be involved in both core promoter recognition and transcriptional activation.

* This study was supported by grants from the Ministry of Education, Science, Sports and Culture of Japan and from the CREST Japan Science and Technology Corporation, the Uehara Memorial Foundation, the Asahi Glass Foundation, the NAITO Foundation, the Sumitomo Foundation, and the Novartis Foundation (Japan) for the Promotion of Science.The costs of publication of this article were defrayed in part by the payment of page charges. The article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

Dagger To whom correspondence should be addressed: Division of Molecular and Cellular Biology, Science of Biological Supramolecular Systems, Graduate School of Integrated Science, Yokohama City University, 1-7-29, Suehiro-cho, Tsurumi-ku, Yokohama, Kanagawa 230-0045, Japan. Tel.: 045-508-7237; Fax: 045-508-7369; E-mail: kokubo@tsurumi.yokohama-cu.ac.jp.

Copyright © 2001 by The American Society for Biochemistry and Molecular Biology, Inc.
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