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Molecular and Cellular Biology, September 2000, p. 6466-6475, Vol. 20, No. 17
0270-7306/00/$04.00+0

Structure and Dynamic Properties of a Glucocorticoid Receptor-Induced Chromatin Transition

Terace M. Fletcher, Byung-Woo Ryu,dagger Christopher T. Baumann, Barbour S. Warren, Gilberto Fragoso, Sam John,Dagger and Gordon L. Hager*

Laboratory of Receptor Biology and Gene Expression, National Cancer Institute, National Institutes of Health, Bethesda, Maryland 20892-5055

Received 17 March 2000/Returned for modification 24 April 2000/Accepted 7 June 2000

Activation of the mouse mammary tumor virus (MMTV) promoter by the glucocorticoid receptor (GR) is associated with a chromatin structural transition in the B nucleosome region of the viral long terminal repeat (LTR). Recent evidence indicates that this transition extends upstream of the B nucleosome, encompassing a region larger than a single nucleosome (G. Fragoso, W. D. Pennie, S. John, and G. L. Hager, Mol. Cell. Biol. 18:3633-3644). We have reconstituted MMTV LTR DNA into a polynucleosome array using Drosophila embryo extracts. We show binding of purified GR to specific GR elements within a large, multinucleosome array and describe a GR-induced nucleoprotein transition that is dependent on ATP and a HeLa nuclear extract. Previously uncharacterized GR binding sites in the upstream C nucleosome region are involved in the extended region of chromatin remodeling. We also show that GR-dependent chromatin remodeling is a multistep process; in the absence of ATP, GR binds to multiple sites on the chromatin array and prevents restriction enzyme access to recognition sites. Upon addition of ATP, GR induces remodeling and a large increase in access to enzymes sites within the transition region. These findings suggest a dynamic model in which GR first binds to chromatin after ligand activation, recruits a remodeling activity, and is then lost from the template. This model is consistent with the recent description of a "hit-and-run" mechanism for GR action in living cells (J. G. McNally, W. G. Müller, D. Walker, and G. L. Hager, Science 287:1262-1264, 2000).

* Corresponding author. Mailing address: Laboratory of Receptor Biology and Gene Expression, Building 41, B602, 41 Library Dr., National Cancer Institute, National Institutes of Health, Bethesda, MD 20892-5055. Phone: (301) 496-9867. Fax: (301) 496-4951. E-mail: hagerg{at}exchange.nih.gov.

dagger Present address: Department of Oncology, Johns Hopkins University School of Medicine, Baltimore, MD 21205-2196.

Dagger Present address: GeneSoft, Inc., South San Francisco, CA 94080.

Molecular and Cellular Biology, September 2000, p. 6466-6475, Vol. 20, No. 17
0270-7306/00/$04.00+0


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