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J. Biol. Chem., Vol. 277, Issue 5, 3202-3209, February 1, 2002
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§,
From the 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.
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
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.
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