Figure 1.

Results of yeast two-hybrid retest with bovine Nub1. The pGAD10 plasmid containing bovine Nub1 was transformed into yeast strain AH109 with either the pGBKT7–bovine Aipl1 plasmid or the pGBKT7–empty plasmid. Transformants were grown on two different types of plates: (i) plates containing synthetic dropout media lacking Leu and Trp (−L/−W) to confirm transformation and (ii) plates containing dropout media with X-α-galactosidase (X-α-gal) and 3-AT (5 mM) but lacking Leu, Trp, and His (−L/−W/−H/+X-α-gal/+3AT) to assay for activation of the HIS3 and MEL1 reporters. (A) Transformation with pGBKT7–Aipl1 and pGAD10–Nub1 plasmids grown on −L/−W plates demonstrate that the transformation worked. (B) Transformation with pGBKT7–empty and pGAD10–Nub1 plasmids grown on −L/−W plates demonstrate that the transformation was successful. (C) Transformation with pGBKT7–Aipl1 and PGAD10–Nub1 plasmids. Formation of blue colonies on −L/−W/−H/+X-α-gal/+3AT plates indicates activation of MEL1 and HIS3 reporters and therefore the presence of a protein–protein interaction between AIPL1 and NUB1. (D) Transformation with pGBKT7–empty and pGAD10–bovine Nub1 plasmids. Lack of colony formation on −L/−W/−H/+X-α-gal/+3AT plates confirms that NUB1 requires the presence of AIPL1 to activate transcription of the HIS and MEL1 reporters.