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Nucleic Acids Research, 1992, Vol. 20, No. 17 4525-4531
© 1992

MOLECULAR BIOLOGY

Ligand dependence of estrogen receptor induced changes in chromatin structure

David M. Gilbert*, Régine Losson and Pierre Chambon

Laboratoire de Génétique Moléculaire des Eucaryotes, CNRS, Unité 184 de Biologie Moléculaire et de Génie Génétique, INSEAM, Institut de Chimie Biologique Faculté de Médecine, 11 rue Humann, 67085 Strasbourg Cedex, France

*To whom correspondence should be addressed at: Roche Institute of Molecular Biology, 340 Kingsland Street, Nutley, NJ 07110, USA

Received May 27, 1992. Revised July 20, 1992. Accepted July 20, 1992.

To determine whether the human estrogen receptor requires ligand to bind to its cognate estrogen receptor element (ERE) in vivo, we have examined the structure of chromatin at a chromosomally integrated ERE-URA3 reporter gene in yeast, and the influence of ligand bound and ligand free estrogen receptors on that structure. Using indirect end-labelling to map DNasel and micrococcal nuclease sensitive sites, we found that receptor induced alterations in chromatin structure were completely dependent upon the presence of estradiol. These same alterations in chromatin structure were induced by a truncated estrogen receptor with both TAF-1 and TAF-2 transactivation functions deleted, suggesting that DNA binding per se disrupts chromatin structure. These results support models in which the estrogen receptor requires ligand to bind to the ERE in vivo.


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