Figure 2.

Homotypic interaction of RHD3 is altered by mutant forms of RHD3. Split ubiquitin-based interaction assay of wild type RHD3 (#2) and mutant forms of RHD3 (#4, #5 and #6) on hisditine minus medium SC-Leu-Trp-His (right plate). The interaction was confirmed by a β-galactosidase activity assay (right plate). The left plate was cells growing on synthetic complete medium SC-Leu-Trp as mating controls. KAT1-KAT1 (#1)⁴⁷ and RHD3-P24σ1d (#3) are used as controls. Note the altered RHD3 interaction by S51N (#4), RHD3–1(A575V) (#5) and T75A (#6) mutations. The number of “+” indicates the intensity of interactions evaluated by colony growth on SC-Leu-Trp-His vs. SC-Leu-Trp. The positive control KAT1-KAT1 was scored as ++++. The assay was conducted according to Chen et al. (2011).²³