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J. Biol. Chem., Vol. 281, Issue 7, 3936-3942, February 17, 2006

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Mutated Yeast Heat Shock Transcription Factor Activates Transcription Independently of Hyperphosphorylation^*

From the Division of Health Sciences, Graduate School of Medical Science, Kanazawa University, 5-11-80 Kodatsuno, Kanazawa, Ishikawa 920-0942, Japan

The homotrimeric heat shock transcription factor (HSF) binds to the heat shock element of target genes and regulates transcription in response to various stresses. The Hsf1 protein of Saccharomyces cerevisiae is extensively phosphorylated upon heat shock; a modification that is under positive regulation by its C-terminal regulatory domain (CTM). Hyperphosphorylation has been implicated in gene-specific transcriptional activation. Here, we surveyed genes whose heat shock response is reduced by a CTM mutation. The CTM is indispensable for transcription via heat shock elements bound by a single Hsf1 trimer but is dispensable for transcription via heat shock elements bound by Hsf1 trimers in a cooperative manner. Intragenic mutations located within or near the wing region of the winged helix-turn-helix DNA-binding domain suppress the temperature-sensitive growth phenotype associated with the CTM mutation and enable Hsf1 to activate transcription independently of hyperphosphorylation. Deletion of the wing partially restores the transcriptional defects of the unphosphorylated Hsf1. These results demonstrate a functional link between hyperphosphorylation and the wing region and suggest that this modification is involved in a conformational change of a single Hsf1 trimer to an active form.

Received for publication, October 4, 2005 , and in revised form, December 16, 2005.

^* This work was supported in part by grants-in-aid for Scientific Research from the Ministry of Education, Sciences, Sports, and Culture (H. S.). The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

The on-line version of this article (available at http://www.jbc.org) contains supplemental data.

¹ Recipient of a Japan Society of the Promotion of Science fellowship (DC2).

² To whom correspondence should be addressed. Tel.: 81-76-265-2588; Fax: 81-76-234-4369; E-mail: sakurai{at}kenroku.kanazawa-u.ac.jp.

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