mTOR-mediated phosphorylation of VAMP8 and SCFD1 regulates autophagosome maturation

• Published in: Nature communications 2021-11, Vol.12 (1), p.6622-6622, Article 6622
• Main Authors: Huang, Hong, Ouyang, Qinqin, Zhu, Min, Yu, Haijia, Mei, Kunrong, Liu, Rong
• Format: Article
• Language: English
• Subjects: 14/1; 14/19; 631/80/313/2104; 631/80/39; 631/80/642/1624; 64/60; 82/58; 82/80; 82/83; Adaptor Proteins, Vesicular Transport - metabolism; Animals; Autophagosomes - genetics; Autophagosomes - metabolism; Autophagy; Complex formation; Depletion; HEK293 Cells; Humanities and Social Sciences; Humans; Lipids; Lysosomes; Male; Membrane Fusion - physiology; Mice; Mice, Inbred C57BL; multidisciplinary; N-Ethylmaleimide-sensitive protein; Phagocytosis; Phosphorylation; Proteins; Qa-SNARE Proteins - metabolism; Qb-SNARE Proteins - metabolism; Qc-SNARE Proteins - metabolism; R-SNARE Proteins - metabolism; Rapamycin; Recruitment; Science; Science (multidisciplinary); SNAP receptors; TOR protein; TOR Serine-Threonine Kinases - metabolism
• ISSN: 2041-1723; 2041-1723
• DOI: 10.1038/s41467-021-26824-5

The mammalian target of rapamycin (mTORC1) has been shown to regulate autophagy at different steps. However, how mTORC1 regulates the N-ethylmaleimide-sensitive protein receptor (SNARE) complex remains elusive. Here we show that mTORC1 inhibits formation of the SNARE complex (STX17-SNAP29-VAMP8) by phosphorylating VAMP8, thereby blocking autophagosome-lysosome fusion. A VAMP8 phosphorylation mimic mutant is unable to promote autophagosome-lysosome fusion in vitro. Furthermore, we identify SCFD1, a Sec1/Munc18-like protein, that localizes to the autolysosome and is required for SNARE complex formation and autophagosome-lysosome fusion. VAMP8 promotes SCFD1 recruitment to autolysosomes when dephosphorylated. Consistently, phosphorylated VAMP8 or SCFD1 depletion inhibits autophagosome-lysosome fusion, and expression of phosphomimic VAMP8 leads to increased lipid droplet accumulation when expressed in mouse liver. Thus, our study supports that mTORC1-mediated phosphorylation of VAMP8 blocks SCFD1 recruitment, thereby inhibiting STX17-SNAP29-VAMP8 complex formation and autophagosome-lysosome fusion. Autophagy relies on coordinated fusion of organelle membranes, although the interplay between the regulatory machinery is not well studied. Here, the authors show that SNARE complex formation is inhibited by mTORC1 phosphorylation of VAMP8, which prevents autophagosome-lysosome fusion.