Methods to analyze SNARE-dependent vesicular fusion events that regulate autophagosome biogenesis.
Bottom Line: Autophagy is an important catabolic pathway that preserves cellular homeostasis.The formation of autophagosomes is a complex process requiring the reorganization of membranes from different compartments.These two steps are essential for the maturation of small single-membrane autophagic precursors containing ATG16L1 and mATG9 proteins into double-membrane autophagosomes.
Affiliation: Department of Medical Genetics, Cambridge Institute for Medical Research, University of Cambridge, Cambridge CB2 0XY, UK.Show MeSH
Mentions: We observed that ATG16L1 and mATG9 both traffic via the plasma membrane. However, they are located in distinct clathrin-coated pits and are internalized and trafficked through largely different routes . mATG9 follows the transferrin receptor pathway through early endosomes and recycling endosomes, whereas ATG16L1 reaches the recycling endosomes but has negligible association with early endosomes. The two different pools of vesicles carrying mATG9 and ATG16L1 coalesce in the recycling endosomes at a stage prior to phagophore formation . If one inhibits this process by knocking down VAMP3, then autophagosome formation is impaired (Fig. 1).
Affiliation: Department of Medical Genetics, Cambridge Institute for Medical Research, University of Cambridge, Cambridge CB2 0XY, UK.