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PMID: 15099516 Published · ppublish English Journal Article

Rapid periodic binding and displacement of the glucocorticoid receptor during chromatin remodeling.

Molecular cell ·Vol. 14 ·No. 2 ·2004-04-23 ·Pages 163-74

Nagaich AK, Walker DA, Wolford R, Hager GL

Abstract

An ultrafast UV laser crosslinking assay has provided novel insights into the progression of the SWI/SNF-mediated chromatin-remodeling reaction and transcription factor binding in real time. We demonstrate site-specific crosslinking between the glucocorticoid receptor (GR), the hSWI/SNF chromatin-remodeling complex, and the mouse mammary tumor virus (MMTV) promoter assembled in an array of correctly positioned nucleosomes. GR first demonstrates rapid binding to the promoter and then is actively displaced from the template during the remodeling reaction. This displacement reaction requires the hSWI/SNF complex and ATP, is specific to the nucleoprotein template, and is accompanied by a core histone rearrangement. The hSWI/SNF complex associates with random positions on the chromatin template in the absence of GR but is recruited specifically to the B/C region when GR is included. These results indicate that enhancement of hSWI/SNF-mediated factor accessibility, a hallmark of chromatin remodeling, is in some cases transient, reversible, and periodic.

MeSH Terms
Adenosine Triphosphate/metabolism Animals Binding Sites Chromatin/metabolism Cross-Linking Reagents/metabolism DNA/genetics,metabolism DNA Helicases Electrophoresis, Polyacrylamide Gel Formaldehyde/metabolism Gene Expression Regulation, Viral Gene Targeting HeLa Cells Histones/metabolism Humans Lasers Mammary Tumor Virus, Mouse/genetics,metabolism Mice Models, Biological Nuclear Proteins/metabolism Nucleosomes/chemistry,metabolism Point Mutation Precipitin Tests Promoter Regions, Genetic Protein Binding Receptors, Glucocorticoid/metabolism Templates, Genetic Time Factors Transcription Factors/metabolism Ultraviolet Rays
Chemicals
Chromatin Cross-Linking Reagents Histones Nuclear Proteins Nucleosomes Receptors, Glucocorticoid Transcription Factors Formaldehyde Adenosine Triphosphate DNA SMARCA4 protein, human Smarca4 protein, mouse DNA Helicases
Authors & Affiliations
4 authors, click to expand affiliations / ORCID
Nagaich Akhilesh K
Laboratory of Receptor Biology and Gene Expression, Building 41, B602, 41 Library Drive, National Cancer Institute, National Institutes of Health, Bethesda, MD 20892, USA.
Walker Dawn A
Wolford Ron
Hager Gordon L
Article Info
Journal
Molecular cell
Abbr.
Mol Cell
ISSN
1097-2765
Published
2004-04-23
Pages
163-74
Language
English
Region
United States
NLM ID
9802571
Subset
IM
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