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Molecular and Cellular Biology, January 2001, p. 534-547, Vol. 21, No. 2
Laboratory of Cellular and Developmental
Biology (NIDDK), National Institutes of Health, Bethesda, Maryland
20892-2715
Received 3 October 2000/Returned for modification 26 October
2000/Accepted 1 November 2000
The yeast CUP1 gene is activated by the
copper-dependent binding of the transcriptional activator, Ace1p. An
episome containing transcriptionally active or inactive
CUP1 was purified in its native chromatin structure from
yeast cells. The amount of RNA polymerase II on CUP1 in the
purified episomes correlated with its transcriptional activity in vivo.
Chromatin structures were examined by using the monomer extension
technique to map translational positions of nucleosomes. The chromatin
structure of an episome containing inactive CUP1 isolated
from ace1
0270-7306/01/$04.00+0 DOI: 10.1128/MCB.21.2.534-547.2001
Remodeling of Yeast CUP1 Chromatin Involves
Activator-Dependent Repositioning of Nucleosomes over the Entire Gene
and Flanking Sequences
and
cells is organized into clusters of
overlapping nucleosome positions separated by linkers. Novel nucleosome
positions that include the linkers are occupied in the presence of
Ace1p. Repositioning was observed over the entire CUP1 gene
and its flanking regions, possibly over the entire episome. Mutation of
the TATA boxes to prevent transcription did not prevent repositioning,
implicating a chromatin remodeling activity recruited by Ace1p. These
observations provide direct evidence in vivo for the nucleosome sliding
mechanism proposed for remodeling complexes in vitro and indicate that
remodeling is not restricted to the promoter but occurs over a
chromatin domain including CUP1 and its flanking sequences.
*
Corresponding author. Mailing address: Laboratory of
Cellular and Developmental Biology (NIDDK), National Institutes of
Health, Building 6, Room B1-12, Bethesda, MD 20892-2715. Phone: (301) 496-6966. Fax: (301) 496-5239. E-mail:
djclark{at}helix.nih.gov.
Present address: Department of Cell Biology and Human Anatomy,
University of California
Davis, Davis, CA 95616.
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