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Molecular and Cellular Biology, June 2004, p. 4651-4663, Vol. 24, No. 11
0270-7306/04/$08.00+0     DOI: 10.1128/MCB.24.11.4651-4663.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.

Temporal Recruitment of Transcription Factors and SWI/SNF Chromatin-Remodeling Enzymes during Adipogenic Induction of the Peroxisome Proliferator-Activated Receptor {gamma} Nuclear Hormone Receptor

Nunciada Salma,1 Hengyi Xiao,1 Elisabetta Mueller,2 and Anthony N. Imbalzano1*

Department of Cell Biology, University of Massachusetts Medical School, Worcester, Massachusetts 01655,1 Department of Cancer Biology, Dana-Farber Cancer Institute, Boston, Massachusetts 021152

Received 23 September 2003/ Returned for modification 5 December 2003/ Accepted 18 March 2004

The peroxisome proliferator-activated receptor gamma (PPAR{gamma}) regulates adipogenesis, lipid metabolism, and glucose homeostasis, and roles have emerged for this receptor in the pathogenesis and treatment of diabetes, atherosclerosis, and cancer. We report here that induction of the PPAR{gamma} activator and adipogenesis forced by overexpression of adipogenic regulatory proteins is blocked upon expression of dominant-negative BRG1 or hBRM, the ATPase subunits of distinct SWI/SNF chromatin-remodeling enzymes. We demonstrate that histone hyperacetylation and the binding of C/EBP activators, polymerase II (Pol II), and general transcription factors (GTFs) initially occurred at the inducible PPAR{gamma}2 promoter in the absence of SWI/SNF function. However, the polymerase and GTFs were subsequently lost from the promoter in cells expressing dominant-negative SWI/SNF, explaining the inhibition of PPAR{gamma}2 expression. To corroborate these data, we analyzed interactions at the PPAR{gamma}2 promoter in differentiating preadipocytes. Changes in promoter structure, histone hyperacetylation, and binding of C/EBP activators, Pol II, and most GTFs preceded the interaction of SWI/SNF enzymes with the PPAR{gamma}2 promoter. However, transcription of the PPAR{gamma}2 gene occurred only upon subsequent association of SWI/SNF and TFIIH with the promoter. Thus, induction of the PPAR{gamma} nuclear hormone receptor during adipogenesis requires SWI/SNF enzymes to facilitate preinitiation complex function.

* Corresponding author. Mailing address: Department of Cell Biology, University of Massachusetts Medical School, 55 Lake Ave. North, Worcester, MA 01655. Phone: (508) 856-1029. Fax: (508) 856-5612. E-mail: anthony.imbalzano{at}umassmed.edu.

Molecular and Cellular Biology, June 2004, p. 4651-4663, Vol. 24, No. 11
0022-538X/04/$08.00+0     DOI: 10.1128/MCB.24.11.4651-4663.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.



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