FIG. 8.

RING finger mutated SNURF is capable of enhancing AR-dependent transactivation but not basal transcription. (A) Influence of SNURF and SNURF(C→S) on transcription from the AR-dependent probasin promoter. CV-1 cells were transfected with 5 μg of pPB(−285/+32)-LUC reporter, 1 μg of pSG5-rAR, 2 μg of pCMVβ, and 5 μg of expression vectors of SNURF, SNURF(C→S), or SNURFΔID/(C→S) or the empty expression vector (pcDNA-3.1+) in the presence or absence of 100 nM testosterone (T) as indicated. (B) Transcription from a minimal AR-dependent promoter. Experimental conditions were as described for panel A, except that 5 μg of pARE₂-TATA-LUC was used as the reporter. (C) Effect of SNURF and SNURF(C→S) on transcription from a minimal TATA-LUC promoter. Experimental conditions were as described for panel A, except that 5 μg of pTATA-LUC in the absence of pSG5-rAR was used. (D) Influence of SNURF and SNURF(C→S) on transcription from a simple Sp1-AP1-TATA promoter. CV-1 cells were transfected as described above but in the absence of pSG5-rAR, and 5 μg of pSp1-AP1-TATA-LUC containing a Sp1 and an AP1 binding site upstream of TATA sequence was used as the reporter. The amounts of pcDNA-SNURF and pcDNA-SNURF(C→S) are given in micrograms. Luciferase (LUC) activities were normalized for transfection efficiency by using the β-galactosidase activity.