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
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
(RXR
) and the murine peroxisome proliferator activated receptor
(PPAR
). Of the known PPAR
isoforms, PPAR
Is the predominant form expressed In adipose tissue. These results suggest that PPAR
serves a unique function among PPA
family members as an important regulator of adipocyte-specific gene expression.
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