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Originally published In Press as doi:10.1074/jbc.M001409200 on May 2, 2000
J. Biol. Chem., Vol. 275, Issue 30, 22678-22685, July 28, 2000
Transcriptional Regulation of the Yeast PHO8 Promoter
in Comparison to the Coregulated PHO5 Promoter*
Martin
Münsterkötter,
Slobodan
Barbaric , and
Wolfram
Hörz§
From the Adolf-Butenandt-Institut, Molekularbiologie,
Universität München, Schillerstrasse 44, 80336 München, Germany
Expression of the PHO8 and
PHO5 genes that encode a nonspecific alkaline and acid
phosphatase, respectively, is regulated in response to the
Pi concentration in the medium by the same transcription
factors. Upon induction by phosphate starvation, both promoters undergo
characteristic chromatin remodeling, yet the extent of remodeling at
the PHO8 promoter is significantly lower than at
PHO5. Despite the coordinate regulation of the two promoters, the PHO8 promoter is almost 10 times weaker than
PHO5. Here we show that of two Pho4 binding sites that had
been previously mapped at the PHO8 promoter in
vitro, only the high affinity one, UASp2, is functional in
vivo. Activation of the PHO8 promoter is partially
Pho2-dependent. However, unlike at PHO5, Pho4
can bind strongly to its binding site in the absence of Pho2 and
remodel chromatin in a Pho2-independent manner. Replacement of the
inactive UASp1 element by the UASp1 element from the PHO5
promoter results in more extensive chromatin remodeling and a
concomitant 2-fold increase in promoter activity. In contrast,
replacement of the high affinity UASp2 site with the corresponding site
from PHO5 precludes chromatin remodeling completely and as
a consequence promoter activation, despite efficient binding of Pho4 to
this site. Deletion of the promoter region normally covered by
nucleosomes 3 and 2 results in a 2-fold increase in promoter
activity, further supporting a repressive role of these nucleosomes.
These data show that there can be strong binding of Pho4 to a UAS
element without any chromatin remodeling and promoter activation. The close correlation between promoter activity and the extent of chromatin
disruption strongly suggests that the low level of PHO8 induction in comparison with PHO5 is partly due to the
inability of Pho4 to achieve complete chromatin remodeling at this promoter.
*
This work was supported by Deutsche Forschungsgemeinschaft
Grant SFB 190, Fonds der Chemischen Industrie (to W. H.), and by Pliva, Zagreb (to S. B.).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.
Present address: Lab. of Biochemistry, Faculty of Food Technology
and Biotechnology, University of Zagreb, Pierottijeva 6, 10000 Zagreb, Croatia.
§
To whom correspondence should be addressed:
Adolf-Butenandt-Institut, Molekularbiologie, Universität
München, Schillerstr. 44, 80336 München, Germany. Fax:
49-89-5996440; E-mail: hoerz@bio. med.uni-muenchen.de.
Copyright © 2000 by The American Society for Biochemistry and Molecular Biology, Inc.

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Copyright © 2000 by the American Society for Biochemistry and Molecular Biology.
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