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