Figure 7.
Failure of uninducible Gal80p mutants in interaction with HA-Gal3p. Whole-cell extracts were prepared from uninduced yeast cells overexpressing HA-Gal3p and the wild-type or one of the mutant Gal80ps; WT, S0, S1, S2, DE21, and − at top represent wild-type GAL80, GAL80S0, GAL80S1, GAL80S2, GAL80DE21, and vacant vector, respectively. Immunoprecipitation was performed in the presence of 1 mM galactose/2 mM ATP. Precipitates were analyzed by immunoblotting with rabbit anti-Gal80p antibody (A) and rabbit anti-Gal3p antibody (B). Twenty micrograms of protein of each whole-cell extract were subjected to immunoblotting using rabbit anti-Gal80p antibody (C).