Nucleic Acids Research, 1995, Vol. 23, No. 9 1536-1543
© 1995
MOLECULAR BIOLOGY |
Transcriptional activation by the orphan nuclear receptor ARP-1
Department of Cardiovascular Molecular Biology Lederle Laboratories, 401 North Middletown Road, Pearl River, NY 10965, USA
* To whom correspondence should be addressed
Received December 20, 1994. Revised March 13, 1995. Accepted March 13, 1995.
ARP-1 is a ubiquitous orphan nuclear receptor that binds to a site (site A) in the apolipoprotein Al (apoAl) liver-specific enhancer and represses its transcriptional activity In hepatoblastoma HepG2 cells. Electro-phoretic mobility shift analysis of HepG2 cell nuclear extracts showed that in addition to ARP-1, site A also binds the orphan nuclear receptors Ear-2 and HNF-4. In in vitro transcription assays, Hela cell nuclear extracts which contain ARP-1 had no effect on transcription from a basal promoter linked to multiple copies of site A. However, supplementation of these extracts with excess amounts of recomblnant ARP-1 resulted In significant stimulation. Supplementation of the extracts with purified polypeptides representing fusions between the ARP-1 N- or C-terminal domains and the yeast activator GAL4 DNA binding domain also stimulated transcription from a basal promoter linked to multiple GAL4 DNA binding sites. Co-immunopreci-prtatlon assays using ARP-1-selective antibodies revealed specific physical Interactions between ARP-1 and the basal transcription factor TFIIB. We conclude that ARP-1 possesses Intrinsic transcription activation potential which is modulated, at least In part, by the intracellular balance of other nuclear receptors that also bind to its cognate DNA binding site.
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