Figure 1. Yeast and Mammalian Two-Hybrid Analyses Identify RANBP17 as a Binding Partner for E12.

(A) Schematic representation of full-length E12 (E12^1-654), full-length RANBP17 (RANBP17^1-1088), the E12 bait (E12^508-654) and the RANBP17 library prey clone (RANBP17^1-252). (B) The indicated pairwise combinations of the bait (indicated at right) and prey constructs (indicated at top) and corresponding empty control vectors were separately transformed into yeast strains Y187 and AH109, mated and protein-protein interaction was assayed as described in Materials and Methods. DDO, media lacking Leu and Trp; TDO media lacking Leu, Trp and His; X-α-Gal, X-α-galactosidase. (C) Mammalian two hybrid analysis. HeLa cells were transfected with the indicated expression constructs for E12^508-654 and RANBP17^1-252 in theVP16 activation domain fusion or the pM binding domain fusions vectors, respectively. Luciferase activities were measured at 48 h post-transfection. Data shown is mean ± S.D., *, p<0.01 for the E12 and RANBP17 co-transfected samples vs. all others shown. Data is representative of three wholly independently conducted experiments, with transfections carried out in triplicate for each such independent experiment.