Figure 5. The dimerization domain of Atg17 regulates Atg17-function.

(a) α-myc immunoblots from co-floatation experiments of Atg9–PLs with recombinant myc-tagged, monomeric Atg17 (Atg17^mono), Atg17^monoTC and Atg1^monoPC. Atg17^mono is strongly inhibited by Atg31–Atg29, but Atg17^monoTC is not activated by Atg1–Atg13. Similar experiments with large unilamellar vesicles (LUVs) lacking Atg9^core served as controls. Input corresponds to 10% of total protein used for co-floatation. (b) Pho8Δ60 assay of wildtype (WT), atg17^mono, and atg17Δ cell lysates to assess autophagic activity under vegetative (grey bars) and starvation (black bars) conditions. Mean values±s.d. of N=3 independent experiments are shown. (c) Co-localization of GFP-Atg8 with mCherry-tagged Atg17 or Atg17^mono under vegetative or autophagy-induced conditions. Atg17^mono is not forming puncta under autophagy-induced conditions. Consequently, autophagosomes are not being formed as apparent from the lack of Atg8-puncta. Scale bar, 2 μm.