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J. Biol. Chem., Vol. 282, Issue 38, 27610-27621, September 21, 2007

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Redundancy of Chromatin Remodeling Pathways for the Induction of the Yeast PHO5 Promoter in Vivo^*

Slobodan Barbaric¹, Tim Luckenbach¹, Andrea Schmid, Dorothea Blaschke, Wolfram Hörz, and Philipp Korber²

From the Adolf-Butenandt-Institut, Universität München, 44 Schillerstrasse, 80336 München, Germany and the Laboratory of Biochemistry, Faculty of Food Technology and Biotechnology, University of Zagreb, 10000 Zagreb, Croatia

Induction of the yeast PHO5 and PHO8 genes leads to a prominent chromatin transition at their promoter regions as a prerequisite for transcription activation. Although induction of PHO8 is strictly dependent on Snf2 and Gcn5, there is no chromatin remodeler identified so far that would be essential for the opening of PHO5 promoter chromatin. Nonetheless, the nonessential but significant involvement of cofactors can be identified if the chromatin opening kinetics are delayed in the respective mutants. Using this approach, we have tested individually all 15 viable Snf2 type ATPase deletion mutants for their effect on PHO5 promoter induction and opening. Only the absence of Snf2 and Ino80 showed a strong delay in chromatin remodeling kinetics. The snf2 ino80 double mutation had a synthetic kinetic effect but eventually still allowed strong PHO5 induction. The same was true for the snf2 gcn5 and ino80 gcn5 double mutants. This strongly suggests a complex network of redundant and mutually independent parallel pathways that lead to the remodeling of the PHO5 promoter. Further, chromatin remodeling kinetics at a transcriptionally inactive TATA box-mutated PHO5 promoter were affected neither under wild type conditions nor in the absence of Snf2 or Gcn5. This demonstrates the complete independence of promoter chromatin opening from downstream PHO5 transcription processes. Finally, the histone variant Htz1 has no prominent role for the kinetics of PHO5 promoter chromatin remodeling.

Received for publication, January 22, 2007 , and in revised form, June 21, 2007.

This paper is dedicated to Gerda and Reiner Luckenbach.

^* This work was supported by the Deutsche Forschungsgemeinschaft (Transregio 5), the 6th Framework Programme of the European Union (Epigenome Network of Excellence), and Ministry of Education, Science, and Technology of the Republic of Croatia Grant 58025 (to S. B.). 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.

¹ These authors contributed equally to this work.

Died Nov. 13, 2005.

² To whom correspondence should be addressed. Tel.: 49-89-218075435; Fax: 49-89-218075425; E-mail: pkorber{at}lmu.de.

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