Fig. 3.

Rim15 promotes Ume6 phosphorylation and functions as a positive regulator of Atg8 induction. (A) Rim15 is required for Ume6 phosphorylation in starvation conditions. Wild-type (WT, BY4742) and rim15Δ cells were grown in rich medium and starved in SD-N for up to 1 h. Cells were collected, and protein extracts were analyzed with anti-Ume6 and anti-Pgk1 (loading control) antisera. (B) Wild-type (WT, YZD005), rim15Δ (YZD006), and rim15Δ ume6Δ (YZD007) cells in a pep4Δ background were grown in rich medium and shifted to SD-N for starvation. Cells were collected at the indicated time points and subjected to immunoblotting with anti-Atg8 and anti-Pgk1 antisera.