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RA-induced RARα degradation is important for optimal RARα-mediated transactivation
RA induces the degradation of its receptor RARα[15] and this is thought to be a resetting mechanism for efficient RA target gene atrial natriuretic peptides [16]. Therefore, we considered it possible that Ski may inhibit RA signaling by influencing this degradation. We first demonstrated that in our system RA did increase RARα turnover.
Fig. 1.
RA-induced RARα degradation is important for optimal RARα-mediated transactivation. (A) COS-1 cells, transfected with plasmid encoding Flag-RARα, were treated without or with 1 μM RA for different times in the presence of CHX as indicated. (B) COS-1 cells were transfected with plasmid encoding Flag-RARα. Twenty-four hours after transfection, cells were treated without or with RA for 8 h in the absence or presence of 5 μM MG132 as indicated. (C) QT6 cells were transfected with CRBPII-Luc, Flag-RARα, and RXR, and after 24 h were left untreated or treated with RA (1 μM) for 12 h. MG132 (5 μM) treatment is as indicated. RLU, relative light units (arbitrary activity). (D) QT6 cells were transfected with Tk-luc reporter and 24 h after transfection the cells were treated with or without MG132 (5 μM) for 12 h. The results are expressed as means ± SD from three independent experiments.