Figure 2.

Summary of the reciprocal binding between members of the YscX and YscY protein families. Protein–protein interactions were determined using the yeast two-hybrid assay. YscX family members were fused to the GAL4 activation domain (in pGADT7), whereas YscY family members were fused to the GAL4 DNA binding domain (in pGBKT7). Pairwise transformations were performed in S. cerevisiae AH109 (Clontech Laboratories) that contained the HIS3 and ADE2 reporter genes. Strength of interactions were determined by the extent of growth on minimal medium devoid of histidine or adenine and recorded after day 4. Green shade indicates robust yeast growth (strong binary interaction), yellow shade reflects modest growth (moderate interaction) and gray shade specifies no growth (no interaction). Due to an intrinsic leakiness with the HIS3 reporter, 4 mM 3-aminotriazole was added to histidine dropout media to suppress false positives (James et al., 1996).