Fig. 5.

The interaction between Atg1 and activated Ypt1 is enhanced in rapamycin-treated cells. (A) Atg1–HA was integrated into wild-type or trs85Δ cells. (Left) Lysed spheroplasts were treated with or without rapamycin, immunoprecipitated with anti-HA antibody, and immunoblotted. (Right) Lysates were immunoblotted with anti-Ypt1 antibody. (B) Atg1–HA preferentially coprecipitates with the GTP form of Ypt1. (Left) Cells expressing the GTP (Q67L) or GDP (S22N) form of Ypt1 were converted to spheroplasts, treated with rapamycin, and processed as in A. (Right) Lysates were immunoblotted with anti-Ypt1 antibody.