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Originally published In Press as doi:10.1074/jbc.M209250200 on October 14, 2002

J. Biol. Chem., Vol. 278, Issue 1, 18-26, January 3, 2003
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A Novel Archaeal Transcriptional Regulator of Heat Shock Response*

Gudrun Vierke, Afra Engelmann, Carina Hebbeln, and Michael ThommDagger

From the Institut für Allgemeine Mikrobiologie, Am Botanischen Garten 1-9, 24118 Kiel, Germany

Archaea have a eukaryotic type of transcriptional machinery containing homologues of the transcription factors TATA-binding protein (TBP) and TFIIB (TFB) and a pol II type of RNA polymerase, whereas transcriptional regulators identified in archaeal genomes have bacterial counterparts. We describe here a novel regulator of heat shock response, Phr, from the hyperthermophilic archaeon Pyrococcus furiosus that is conserved among Euryarchaeota. The protein specifically inhibited cell-free transcription of its own gene and from promoters of a small heat shock protein, Hsp20, and of an AAA+ ATPase. Inhibition of transcription was brought about by abrogating RNA polymerase recruitment to the TBP/TFB promoter complex. Phr bound to a 29-bp DNA sequence overlapping the transcription start site. Three sequences conserved in the binding sites of Phr, TTTA at -10, TGGTAA at the transcription start site, and AAAA at position +10, were required for Phr binding and are proposed as consensus regulatory sequences of Pyrococcus heat shock promoters. Shifting the growth temperature from 95 to 103 °C caused a dramatic increase of mRNA levels for the aaa+ atpase and phr genes, but expression of the Phr protein was only weakly stimulated. Our findings suggest that heat shock response in Archaea is negatively regulated by a mechanism involving binding of Phr to conserved sequences.

* This work was supported by a grant from the Deutsche Forschungsgemeinschaft priority program and from the Fonds der Chemischen Industrie (to M. T.).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: Universität Regensburg, Universitätsstr. 31, 93040 Regensburg, Germany. Tel.: 49-941-943-3160; Fax: 49-941-943-2403; E-mail: mthomm@ifam.uni-kiel.de or Michael.Thomm{at}Biologie.Uni-Regensburg.de.

Copyright © 2003 by The American Society for Biochemistry and Molecular Biology, Inc.
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