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ATG13 dynamics in nonselective autophagy and mitophagy: insights from live imaging studies and mathematical modeling

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journal contribution
posted on 22.04.2020, 16:28 by Piero Dalle Pezze, Eleftherios Karanasios, Varvara Kandia, Maria Manifava, Simon A. Walker, Nicolas Gambardella Le Novère, Nicholas T. Ktistakis

During macroautophagy/autophagy, the ULK complex nucleates autophagic precursors, which give rise to autophagosomes. We analyzed, by live imaging and mathematical modeling, the translocation of ATG13 (part of the ULK complex) to the autophagic puncta in starvation-induced autophagy and ivermectin-induced mitophagy. In nonselective autophagy, the intensity and duration of ATG13 translocation approximated a normal distribution, whereas wortmannin reduced this effect and shifted to a log-normal distribution. During mitophagy, multiple translocations of ATG13 with increasing time between peaks were observed. We hypothesized that these multiple translocations arise because the engulfment of mitochondrial fragments required successive nucleation of phagophores on the same target, and a mathematical model based on this idea reproduced the oscillatory behavior. Significantly, model and experimental data were also in agreement that the number of ATG13 translocations is directly proportional to the diameter of the targeted mitochondrial fragments. Thus, our data provide novel insights into the early dynamics of selective and nonselective autophagy.

Abbreviations: ATG: autophagy related 13; CFP: cyan fluorescent protein; dsRED: Discosoma red fluorescent protein; GABARAP: GABA type A receptor-associated protein; GFP: green fluorescent protein; IVM: ivermectin; MAP1LC3/LC3: microtubule-associated protein 1 light chain 3; MTORC1: mechanistic target of rapamycin kinase complex 1; PIK3C3/VPS34: phosphatidylinositol 3-kinase catalytic subunit type 3; PtdIns3P: PtdIns-3-phosphate; ULK: unc-51 like autophagy activating kinase.

Funding

This work was funded by the Biotechnology and Biological Sciences Research Council (grant numbers: BBS/E/B/000C0419 and BBS/E/B/000C0434 to NLN; BB/K019155/1 to NTK); and a science policy committee from the Babraham Institute, Cambridge (grant number 2301-R1600-C0341 to NTK).

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