Figure 4.
Regulation of zinc starvation-induced autophagy by TORC1. A–D, phosphorylation status of Atg13 and Sch9 under zinc starvation conditions. A, cells expressing Atg13 tagged with 6×HA were cultured in SD(−zinc). At the indicated time points, lysates were prepared, and the migration of Atg13 was monitored by Western blotting using an anti-HA antibody. B, lysates of cells cultured in SD(−zinc) for 0 or 15 h were incubated with or without λ-protein phosphatases (PPase). C, zinc-starved cells (−zinc 15 h) were supplied with 2 μm zinc for 5 or 10 min in the presence or absence of cycloheximide (CHX) (10 μg/ml). D, cells expressing Sch9 tagged with 6×HA were analyzed as in C. E, processing of GFP-Atg8 in atg11Δ cells expressing a Tor1 hyperactive mutant (TOR1L2134M) or Tor1WT under zinc starvation for 0, 14, and 24 h.