Abstract
Autophagy involves rapid growth of phagophores through membrane addition. Newly added membranes are derived from other organelles through the formation of vesicles carrying the Atg9 protein. Membrane is delivered by fusing these vesicles with the phagophores. Atg9 is, nevertheless, not incorporated in autophagosomes. We now show that this protein is retrieved from phagophores by fission utilizing Dynamin-2 (Dnm2) as the membrane scission protein. Blocking Atg9 recycling by interfering with Dnm2 helps retain Atg9 in autophagosomes and degrades this protein by autophagy. Dnm2 colocalizes with the BAR domain protein Endophilin-B1 (EndoB1/Bif-1) when autophagy is induced, suggestive of transient interactions, but mutations in these proteins have very different effects from which we conclude that they act at different stages of autophagy. Our results show that Dnm2 promotes Atg9 retrieval, while EndoB1 may anchor the autophagy machinery. These data provide novel insights into the roles of membrane scission proteins during autophagy.
