Ctl1 is required for autophagic transport and normal PAS organization.

(A) The predicted membrane topology of Ctl1. (B,C) A fluorescence loss in photobleaching (FLIP) assay revealed the immobilized and non-diffusible pools of Tdh1-YFP. Yellow dots mark the sites of photobleaching. (B) In non-starved cells, only nuclear YFP signal remained in post-FLIP images. (C) In starved cells, vacuolar YFP signal was observed in post-FLIP images of wild-type, but not atg5Δ or ctl1Δ cells. (D) Ring-shaped and C-shaped structures labeled by CFP-Atg8 appeared in ctl1Δ cells after prolonged starvation. (E) Time-lapse images of a ctl1Δ cell. (F) The localization patterns of other Atg proteins in ctl1Δ cells containing CFP-Atg8-labeled structures. (G) Time-lapse images of a ctl1Δ cell expressing both CFP-Atg8 and YFP-Atg2. Bars, 3 µm.