PRIC295, a Nuclear Receptor Coactivator, Identified from PPAR-Interacting Cofactor Complex
Figure 4
PRIC295 interactions with nuclear receptors. (a) In vitro interaction of radiolabeled full-length PRIC295 with nuclear receptors PPAR, RXR, PPAR, TR, CAR, and ER. PRIC295 was radiolabeled using 35S-methionine (Perkin Elmer) and incubated with each shown receptor fusion protein in GST-pulldown assay. () minus indicates in vitro binding interaction in the absence of cognate ligand and (+) plus indicates the presence of receptor-specific ligand during the in vitro binding interaction. Ligands used: Wy-14,643 for PPAR; 9-cis-retinoic acid for RXR; rosiglitazone for PPAR; triiodothyronine (T3)TR; 1,4-Bis[2-(3,5-dichloropyridyloxy)]benzene (TCPOBOP) for CAR; 17--estradiol for ER. All ligands were used at a concentration of 100 M. (b) Interaction of radiolabeled PRIC295 fragments Δ (F1), Δ (F2), and Δ (F3) with GST-fusion nuclear receptor proteins. Loading for each lane is indicated at the base of each panel. Lane 1 input; lane 2 GST alone without ligand; lane 3 GST alone with receptor-specific ligand; lane 4 GST-receptor fusion in the absence ligand; lane 5 in the presence of receptor-specific ligand. All ligands used are as previously described in (a).
