ATP-Dependent Mobilization of the Glucocorticoid Receptor during Chromatin Remodeling

ABSTRACT 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.

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[93]  J. Workman,et al.  Alteration of nucleosome structure as a mechanism of transcriptional regulation. , 1998, Annual review of biochemistry.

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[108]  D. O. Peterson,et al.  Functional elements of the steroid hormone-responsive promoter of mouse mammary tumor virus , 1990, Journal of virology.

[109]  H. Diggelmann,et al.  Dual function of a nuclear factor I binding site in MMTV transcription regulation. , 1989, Nucleic acids research.

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[112]  H. Ponta,et al.  DNA sequences outside the receptor‐binding sites differently modulate the responsiveness of the mouse mammary tumour virus promoter to various steroid hormones. , 1988, The EMBO journal.

[113]  H. Richard-Foy,et al.  Mechanism of dexamethasone 21-mesylate antiglucocorticoid action: II. Receptor-antiglucocorticoid complexes do not interact productively with mouse mammary tumor virus long terminal repeat chromatin. , 1987, Molecular endocrinology.

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[115]  G. Hager,et al.  Steroid-dependent interaction of transcription factors with the inducible promoter of mouse mammary tumor virus in vivo , 1987, Cell.

[116]  M. M. Bradford A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. , 1976, Analytical biochemistry.