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Originally published In Press as doi:10.1074/jbc.M203898200 on May 23, 2002

J. Biol. Chem., Vol. 277, Issue 31, 28247-28255, August 2, 2002
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Glucocorticoid Receptor Domain Requirements for Chromatin Remodeling and Transcriptional Activation of the Mouse Mammary Tumor Virus Promoter in Different Nucleoprotein Contexts*

Erika Krasnickas KeetonDagger §, Terace M. Fletcher§, Christopher T. Baumann§, Gordon L. Hager§, and Catharine L. Smith§||

From the Dagger  Department of Genetics, George Washington University, Washington, D. C. and the § Laboratory of Receptor Biology and Gene Expression, Center for Cancer Research, NCI, National Institutes of Health, Bethesda, Maryland 20852

The glucocorticoid receptor (GR) contains several activation domains, tau 1 (AF-1), tau 2, and AF-2, which were initially defined using transiently transfected reporter constructs. Using domain mutations in the context of full-length GR, this study defines those domains required for activation of the mouse mammary tumor virus (MMTV) promoter in two distinct nucleoprotein configurations. A transiently transfected MMTV template with a disorganized, accessible chromatin structure was largely dependent on the AF-2 domain for activation. In contrast, activation of an MMTV template in organized, replicated chromatin requires both domains but has a relatively larger dependence on the tau 1 domain. Domain requirements for GR-induced chromatin remodeling of the latter template were also investigated. Mutation of the AF-2 helix 12 domain partially inhibits the induction of nuclease hypersensitivity, but the inhibition was relieved in the absence of tau 1, suggesting the occurrence of an important interaction between the two domains. Further mutational analysis indicates that GR-induced chromatin remodeling requires the ligand-binding domain in the region of helix 3. Our study shows that the GR activation surfaces required for transcriptional modulation of a target promoter were determined in part by its chromatin structure. Within a particular cellular environment the GR appears to possess a significant degree of versatility in the mechanism by which it activates a target promoter.


* The costs of publication of this article were defrayed in part by the payment of page charges. The article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

Present address: Chroma Technology Corp., 74 Cotton Mill Hill, Unit A-9, Brattleboro, VT 05301.

|| To whom correspondence should be addressed: National Institutes of Health, Laboratory of Receptor Biology and Gene Expression, Bldg. 41, Rm. B608, 41 Library Dr. MSC 5055, Bethesda, MD 20892-5055. Tel.: 301-496-7538; Fax: 301-496-4951; E-mail: smithcat@exchange.nih.gov.


Copyright © 2002 by The American Society for Biochemistry and Molecular Biology, Inc.
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