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Nucleic Acids Research, 1994, Vol. 22, No. 25 5628-5634
© 1994


Articles

Adipocyte-specific transcription factor ARF6 is a heterodimeric complex of two nuclear hormone receptors, PPAR7 and RXRa

Peter Tontonoz, Reed A. Graves+,, Adriane I. Budavari, Hediye Erdjument-Bromage1, Mary Lui1, Erding Hu, Paul Tempst1 and Bruce M. Spiegelman*,

Dana-Farber Cancer Institute and the Department of Cell Biology, Harvard Medical School Boston, MA 02115 1Molecular Biology Program, Memorial Sloan-Kettering Cancer Center New York, NY 10021, USA

*To whom correspondence should be addressed

+Present address: Department of Medicine. University of Chicago. Chicago. IL 60637, USA

Received September 6, 1994. Revised November 12, 1994. Accepted November 12, 1994.

Previously, we Identified a novel transcription factor, ARF6, as a key regulator of the tissue-specific adipocyte P2 (aP2) enhancer. In order to identify the proteins which comprise the adipocyte ARF6 complex, we have purified this DNA binding activity from a cultured adipocyte cell line. We have developed a system for growth and differentiation of HIB-1B brown adlpocytes in suspension culture that facilitates the production of large quantities of adipocyte nuclear extract. ARF6 was purified from HIB-1B nuclear extract by a combination of conventional and sequence-specific DNA affinity chromotography. Chemical sequencing and mass spectral analysis of tryptlc peptldes derived from the purified polypeptides Identifies the ARF6 complex as a heterodlmer of the retlnold X receptor {alpha} (RXR{alpha}) and the murine peroxisome proliferator activated receptor {gamma} (PPAR{gamma}). Of the known PPAR{gamma} isoforms, PPAR{gamma} Is the predominant form expressed In adipose tissue. These results suggest that PPAR{gamma} serves a unique function among PPA{gamma} family members as an important regulator of adipocyte-specific gene expression.


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