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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
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Transcriptional Regulation of the Yeast PHO8 Promoter in Comparison to the Coregulated PHO5 Promoter*

Martin Münsterkötter, Slobodan BarbaricDagger , 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.

Dagger 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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