FIGURE 3.

Gcn2 activation inhibits TORC1 activity. A, wild type (Y661) cells expressing SCH9-HA together with a galactose-inducible GCN2(E803V) (+) or control vector (−) grown in medium containing 2% glucose were treated with rapamycin (Rap+) or vehicle control (−). TORC1-dependent phosphorylation of Sch9-HA was examined by Western blotting. B, wild type (Y661) cells expressing GFP-ATG8 together with a control vector or vector expressing a galactose-inducible GCN2(E803V) or gcn2(K628R) were grown in the presence of 2% galactose (Gal) for the indicated times. Autophagic cleavage of GFP-Atg8 was assayed by Western blotting. Tpd3 levels were used as loading controls. The experiments were repeated three times, and representative data are shown.