Journal Article

Atg9 is a lipid scramblase that mediates autophagosomal membrane expansion

Tsuji Takuma, Matoba Kazuaki, Noda, Nobuo N, Kotani Tetsuya, Sugita Yuji, Iwata So, Noshiro Daisuke, Tsutsumi Akihisa, Mori Takaharu, Nakatogawa Hitoshi, Kikkawa Masahide, Ohsumi Yoshinori, Fujimoto Toyoshi, Nomura Norimichi

2020

Overview

The molecular function of Atg9, the sole transmembrane protein in the autophagosome-forming machinery, remains unknown. Atg9 colocalizes with Atg2 at the expanding edge of the isolation membrane (IM), where Atg2 receives phospholipids from the endoplasmic reticulum (ER). Here we report that yeast and human Atg9 are lipid scramblases that translocate phospholipids between outer and inner leaflets of liposomes in vitro. Cryo-EM of fission yeast Atg9 reveals a homotrimer, with two connected pores forming a path between the two membrane leaflets: one pore, located at a protomer, opens laterally to the cytoplasmic leaflet; the other, at the trimer center, traverses the membrane vertically. Mutation of residues lining the pores impaired IM expansion and autophagy activity in yeast and abolished Atg9’s ability to transport phospholipids between liposome leaflets. These results suggest that phospholipids delivered by Atg2 are translocated from the cytoplasmic to the luminal leaflet by Atg9, thereby driving autophagosomal membrane expansion.Cryo-EM and liposome assays reveal that Atg9 functions as a lipid scramblase, transporting phospholipids between inner and outer liposome leaflets. Analyses of mutants in yeast support a role for this activity in autophagy.

• Publisher: Nature Publishing Group
• Subject: Autophagy / Endoplasmic reticulum / Lipids / Liposomes / Membranes / Mutation / Phagocytosis / Phospholipids / Pores / Trimers / Yeast