Nucleic Acids Research, Vol 27, Issue 16 3364-3370, Copyright © 1999 by Oxford University Press
DE Cryderman, H Tang, C Bell, DS Gilmour and LL Wallrath
In a variety of organisms, genes placed near heterochromatin are
transcriptionally silenced. In order to understand the molecular mechanisms
responsible for this block in transcription, high resolution in vivo
chromatin structure analysis was performed on two heat shock genes, hsp26
and hsp70. These genes normally reside in euchromatin where GAGA factor and
RNA Pol II are present on the promoter prior to heat shock induction.
P-element transformation experiments led to the identification of stocks in
which these two genes were inserted within heterochromatin of the
chromosome 4 telomeric region. These transgenes exhibit silencing that is
partially suppressed by mutations in the gene encoding HP1. Micrococcal
nuclease analysis revealed that the heterochromatic transgenes were
packaged in a more regular nucleosome array than when located in
euchromatin. High resolution DNase I analysis demonstrated that GAGA factor
and TFIID were not associated with these promoters in heterochromatin;
potassium permanganate experiments showed a loss of Pol II association.
Taken together, these data suggest that occlusion of trans-acting factors
from their cis- acting regulatory elements leading to a block in promoter
potentiation is a mechanism for heterochromatin gene silencing.
ARTICLES
Heterochromatic silencing of Drosophila heat shock genes acts at the level of promoter potentiation
Department of Biochemistry, 4-772 Bowen Science Building, University of Iowa, Iowa City, IA 52242, USA.
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