Retinoid receptors interact with the DRIP/TRAP complex in APL NB4 cells.
(A) Comparison of the protein interactions to retinoid receptor heterodimer with those of the vitamin D3 receptor. Immobilized GST-VDR LBD or GST-RXR LBD/RAR LBD was incubated with 1.2 mg NB4 nuclear extract in the absence (−) or presence (+) of their respective ligands, 1,25-dihydroxy vitamin D3 (lane 2) and t-RA (lane 4). (B) Immunoblot analyses with 2 specific anti-DRIP antibodies. The RXR/RAR-interacting proteins were separated by SDS-PAGE, transferred to a nitrocellulose membrane, and subjected to Western blotting using antibodies to DRIP130 and DRIP150, respectively. (C) In vitro interaction of the DRIP205 protein to RXR/RAR heterodimer or VDR. Purified GST or GST-RXR LBD/RAR LBD and GST-VDR LBD were incubated with in vitro translated 35S-labeled DRIP205 in the absence (−) and presence of 1 μmol/L or 10 μmol/L t-RA (lanes 4, 5) and 1,25-dihydroxy vitamin D3 (lanes 7, 8) respectively. Twenty percent of the DRIP205 protein used was shown as input.
