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Molecular and Cellular Biology, May 2002, p. 3255-3263, Vol. 22, No. 10
0270-7306/02/$04.00+0     DOI: 10.1128/MCB.22.10.3255-3263.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.

ATP-Dependent Mobilization of the Glucocorticoid Receptor during Chromatin Remodeling

Terace M. Fletcher, Nianqing Xiao, Gisele Mautino, Christopher T. Baumann, Ronald Wolford, Barbour S. Warren,,{dagger} and Gordon L. Hager*

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

Received 14 September 2001/ Returned for modification 5 November 2001/ Accepted 5 February 2002

Chromatin remodeling by the glucocorticoid receptor (GR) is associated with activation of transcription at the mouse mammary tumor virus (MMTV) promoter. We reconstituted this nucleoprotein transition with chromatin assembled on MMTV DNA. The remodeling event was ATP dependent and required either a nuclear extract from HeLa cells or purified human Swi/Snf. Through the use of a direct interaction assay (magnetic bead pull-down), we demonstrated recruitment of human Swi/Snf to MMTV chromatin by GR. Unexpectedly, we found that GR is actively displaced from the chromatin template during the remodeling process. ATP-dependent GR displacement was reversed by the addition of apyrase and was specific to chromatin templates. The disengagement reaction could also be induced with purified human Swi/Snf. Although GR apparently dissociated during chromatin remodeling by Swi/Snf, it participated in binding of the secondary transcription factor, nuclear factor 1. These results are paralleled by a recent discovery that the hormone-occupied receptor undergoes rapid exchange between chromatin and the nucleoplasmic compartment in living cells. Both the in vitro and in vivo results are consistent with a dynamic model (hit and run) in which GR first binds to chromatin after ligand activation, recruits a remodeling activity, facilitates transcription factor binding, and is simultaneously lost from the template.

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

{dagger} Present address: Center for the Environment, Cornell University, Ithaca, NY 14850.

Molecular and Cellular Biology, May 2002, p. 3255-3263, Vol. 22, No. 10
0022-538X/02/$04.00+0     DOI: 10.1128/MCB.22.10.3255-3263.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.



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