Phosphomimetic mutation at T109 inhibits the oligomerization of She2 and its interaction with cofactors Srp1 and She3. (A) Yeast two-hybrid assay to detect She2–She2 interaction. Homo- or hetero-oligomerization of wild-type She2 (WT), She2 T109A (T109A), or She2 T109D (T109D) was quantified by measuring β-galactosidase activity (N = 3). (**) P < 0.01; (***) P < 0.005. (B) Yeast two-hybrid assay to detect the interaction between She3 and She2 mutants at T109. Interaction between She3 and wild-type She2 (WT), She2 T109A (T109A), or She2 T109D (T109D) was quantified by measuring β-galactosidase activity (N = 3). (**) P < 0.01. (C) Yeast two-hybrid assay to detect the interaction between Srp1 and She2 mutants at T109. Interaction between Srp1 and wild-type She2 (WT), She2 T109A (T109A), or She2 T109D (T109D) was quantified by measuring β-galactosidase activity (N = 3). (****) P < 0.001.
