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Originally published In Press as doi:10.1074/jbc.M108962200 on November 8, 2001

J. Biol. Chem., Vol. 277, Issue 5, 3202-3209, February 1, 2002
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Neither Reb1p nor Poly(dA·dT) Elements Are Responsible for the Highly Specific Chromatin Organization at the ILV1 Promoter*

José M. A. MoreiraDagger §, Wolfram Hörz, and Steen HolmbergDagger ||

From the Dagger  Department of Genetics, Institute of Molecular Biology, University of Copenhagen, Øster Farimagsgade 2A, DK-1353, Copenhagen K, Denmark and the  Institut für Physiologische Chemie der Universität, München, Schillerstr, 44, 80336 München, Germany

Analysis of the chromatin structure at the yeast ILV1 locus revealed highly positioned nucleosomes covering the entire locus except for a hypersensitive site in the promoter region. All previously identified cis-acting elements required for GCN4-independent ILV1 basal level transcription, including a binding site for the REB1 protein (Reb1p), and a poly(dA·dT) element (26 As out of 32 nucleotides) situated 15 base pairs downstream of the Reb1p-binding site, reside within this hypersensitive site. The existence of a second A·T-rich element (25 As out of 33 nucleotides) present six base pairs upstream of the Reb1p-binding site, suggested that nucleosome exclusion from the hypersensitive site in the ILV1 promoter region might be dictated by synergistic action of the two poly(dA·dT) elements. Replacing one or both of them had, however, no effect on the chromatin structure of the ILV1 promoter, although drastically reduced basal transcription. Similarly, deletion of the Reb1p-binding site, albeit affecting ILV1 expression, had no detectable effect on chromatin at the ILV1 promoter. The absence of a good correlation between effects of these elements on gene activity and on chromatin structure at the ILV1 promoter indicates that the chromatin organization present at the ILV1 promoter is independent of the known regulatory elements and most likely dictated directly by the DNA sequence.

* This work was supported by Junta Nacional de Investigação Científica e Tecnológica Grant BD-2323-IF to J. M. A. M., and grants from the Danish Research Councils, the Novo-Nordisc Foundation, Løvens Kemiske Fabriks Fond, and Lundbeckfonden.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.

§ Current address: Active Biotech Research AB, P.O. Box 724, SE-22007 Lund, Sweden.

|| To whom correspondence should be addressed: Dept. of Genetics, Institute of Molecular Biology, Univ. of Copenhagen, Øster Farimagsgade 2A, DK-1353, Copenhagen K, Denmark. Tel.: 45-3532-2119; Fax: 45-3532-2113; E-mail: gensteen@biobase.dk.

Copyright © 2002 by The American Society for Biochemistry and Molecular Biology, Inc.
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