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Molecular and Cellular Biology, November 1998, p. 6745-6755, Vol. 18, No. 11
Department of Pharmacology, University of
Minnesota Medical School, Minneapolis, Minnesota 55455
Received 12 March 1998/Returned for modification 12 June
1998/Accepted 14 August 1998
The mouse homologue of the human receptor-interacting protein 140 (RIP140) was isolated from a mouse embryonic cDNA library in yeast
two-hybrid screening experiments by using the ligand binding domain
(LBD) of nuclear orphan receptor TR2 as the bait. The
receptor-interacting domains of mouse RIP140 were mapped to the regions
containing the LXXLL motif (where L is leucine and X is any amino
acid), and the RIP140-interacting domain of TR2 was mapped to its
C-terminal 10- to 20-amino-acid sequence, a putative activation
function 2 (AF-2) region. In a GAL4 reporter system and a reporter
driven by the proximal region of the TR2 promoter, RIP140 functioned as
a corepressor for both a GAL4 DNA binding domain (BD)-TR2 fusion and
the wild-type receptor. When tethered to the BD of GAL4, RIP140 exerted
a trans-repressive effect on the GAL4 reporter. In
addition, RIP140 suppressed the retinoic acid (RA) receptor-mediated RA
induction in a dose-dependent manner. Finally, it was demonstrated that
in the presence of RIP140, a cytosolic, green fluorescent
protein-tagged TR2 LBD translocated into the nucleus, and TR2 and
RIP140 were coimmunoprecipitated from the cell extract, indicating that
the interaction between RIP140 and the LBD of TR2 occurred in vivo. The
potential biological role of RIP140 in TR2-modulated transcriptional
activity is discussed.
0270-7306/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.
Cloning and Characterization of Mouse RIP140, a
Corepressor for Nuclear Orphan Receptor TR2
*
Corresponding author. Mailing address: Dept. of
Pharmacology, University of Minnesota, 3-249 Millard Hall, 435 Delaware
St. SE, Minneapolis, MN 55455. Phone: (612) 625-9402. Fax: (612)
625-8408. E-mail: weixx009{at}maroon.tc.umn.edu.
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