Figure 5.

Atg17–Atg29 and Atg1–Atg13 interactions are indispensable for their recruitment to the PAS. (A) Atg29 mutant (Atg29^F67L and Atg29^R71G) is defective for the ATG11-independent PAS localization of Atg1, Atg13, or Atg17. atg11Δatg29Δ cells expressing GFP-tagged Atg1, Atg13, or Atg17 were transformed with a plasmid expressing either Atg29^wild-type, Atg29^F67L, or Atg29^F71G. Cells were treated with rapamycin for 3 h before observation. (B) Atg13^1-448 does not recruit Atg29 to the PAS in atg11Δ cells. atg11Δatg13Δ cells expressing Atg29-GFP were transformed with a plasmid expressing either Atg13^{wild type} or Atg13^1-448. Cells were observed after rapamycin treatment for 3 h. (C) Atg1 kinase activity is not essential for the PAS targeting of Atg17 and Atg29 in atg11Δ. atg11Δatg1Δ cells expressing Atg17-GFP or Atg29-GFP were transformed with a plasmid expressing either wild-type Atg1 or Atg1 kinase-deficient mutant (Atg1^D211A or Atg1^K54A). Cells were observed after rapamycin-treatment for 3 h. Bars, 5 μm.