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PMID: 2846275 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Specific glucocorticoid receptor binding to DNA reconstituted in a nucleosome.

The EMBO journal ·Vol. 7 ·No. 10 ·1988-10-00 ·Pages 3073-9

Perlmann T, Wrange O

Abstract

We have reconstituted a nucleosome with core histones from rat liver using a restriction fragment containing a sequence from the mouse mammary tumour virus (MTV) long terminal repeat (LTR). This sequence harbours glucocorticoid responsive elements (GREs) which mediate glucocorticoid hormone induction of transcription from the MTV promoter via glucocorticoid receptor (GR) binding. Exonuclease III and DNase I footprinting demonstrated that the reconstituted nucleosome was specifically located between positions -219 and -76. A nucleosome was previously shown to be located at a similar or identical position in the MTV promoter in situ and to be structurally altered upon glucocorticoid hormone induction. We demonstrated, by DNase I footprinting, that GR is able to bind sequence specifically to the DNA in the in vitro assembled nucleosome. No evidence for unfolding of the nucleosome was obtained, but the DNase I footprinting pattern demonstrated GR induced local alterations in the DNA.

MeSH Terms
Animals Base Sequence Cloning, Molecular DNA/metabolism DNA-Binding Proteins/metabolism Deoxyribonuclease I/metabolism In Vitro Techniques Mammary Tumor Virus, Mouse/genetics Molecular Sequence Data Nucleosomes/metabolism Plasmids Rats Receptors, Glucocorticoid/metabolism
Chemicals
DNA-Binding Proteins Nucleosomes Receptors, Glucocorticoid DNA Deoxyribonuclease I
Authors & Affiliations
2 authors, click to expand affiliations / ORCID
Perlmann T
Department of Molecular Genetics, Medical Nobel Institute, Karolinska Institutet, Stockholm, Sweden.
Wrange O
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32 references, click to expand
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Article Info
Journal
The EMBO journal
Abbr.
EMBO J
ISSN
0261-4189
Published
1988-10-00
Pages
3073-9
Language
English
Region
England
NLM ID
8208664
PMCID
PMC454694
Subset
IM
External Links
PubMed source PMC Full Text View journal details
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