Transferrin receptor controls both autophagosome formation and closure via phosphatidylinositol 3-phosphate synthesis.

Autophagosome formation involves multiple sequential steps that need to be coordinated and linked. Here, we describe in mammalian cells that the transferrin receptor (TfR) links LC3 family conjugation to phagophore membranes, an early step in autophagosome biogenesis, with subsequent autophagosome closure. TfR depletion impairs autophagic flux and its overexpression stimulates this catabolic process in an iron-independent manner. TfR is ubiquitinated by the ubiquitin ligase MARCH8 in the RAB11A-LC3B-positive membranes that are conjugated by LC3 family members from which phagophores emanate. Ubiquitinated TfR recruits the VPS34 component VPS15, enabling phosphatidylinositol 3-phosphate (PI(3)P) synthesis on nascent autophagosome membranes. This PI(3)P is not only important for LC3-lipid conjugation but also for subsequent phagophore closure, where TfR-dependent PI(3)P recruits the endosomal sorting complexes required for transport (ESCRT) complex. This TfR activity occurs after endocytosis of iron-containing transferrin, its canonical function, as TfR only binds VPS15 after iron detachment from transferrin that is enabled by pH lowering in the endocytic compartment.