Specific phosphoresidues modulate the capacity of She2 to promote the asymmetric distribution of Ash1. (A) Genetic assay to assess Ash1 asymmetric distribution in She2 mutants. Serial dilutions of K5547 + YCP22-She2-myc, K5547 + YCP22-She2-M5A-myc, K5547 + YCP22-She2-T109A-myc, K5547 + YCP22-She2-T109D-myc, and K5547 + YCP22 empty. (B) Impact of various phospho-null and phosphomimetic mutants of She2 on the growth of K5547 on −ADE −TRP medium. (C) Western blotting of She2-myc WT, T109A, and T109D expression in the K5547 strain. YCP: K5547 + YCPlac22 empty. Tubulin was used as a loading control. (D) Quantification of She2 WT, She2 T109A, and She2 T109A expression in the K5547 strain. (***) P < 0.005. N = 3. (E) Relative expression of SHE2 mRNA quantified by RT-qPCR in K5547 strain expressing YCPlac22 (YCP), She2 WT, She2 T109A, or She2 T109D proteins. N = 2.
