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SNARE proteins are required for macroautophagy
Authors:Nair Usha  Jotwani Anjali  Geng Jiefei  Gammoh Noor  Richerson Diana  Yen Wei-Lien  Griffith Janice  Nag Shanta  Wang Ke  Moss Tyler  Baba Misuzu  McNew James A  Jiang Xuejun  Reggiori Fulvio  Melia Thomas J  Klionsky Daniel J
Institution:1 Life Sciences Institute, University of Michigan, Ann Arbor, MI 48109, USA
2 Science Technology and Research Scholars, Yale College, New Haven, CT 06520, USA
3 Cell Biology Program, Memorial Sloan Kettering Cancer Center, New York, NY 1002, USA
4 Department of Cell Biology, Yale University, New Haven, CT 06520, USA
5 Department of Cell Biology and Institute of Biomembranes, University Medical Centre Utrecht, 3584 CX Utrecht, The Netherlands
6 Department of Biochemistry and Cell Biology, Rice University, Houston, TX 77005, USA
7 Collaborative Open Research Center, Kogakuin University, Hachioji, Tokyo 192-0015, Japan
Abstract:Macroautophagy mediates the degradation of long-lived proteins and organelles via the de novo formation of double-membrane autophagosomes that sequester cytoplasm and deliver it to the vacuole/lysosome; however, relatively little is known about autophagosome biogenesis. Atg8, a phosphatidylethanolamine-conjugated protein, was previously proposed to function in autophagosome membrane expansion, based on the observation that it mediates liposome tethering and hemifusion in vitro. We show here that with physiological concentrations of phosphatidylethanolamine, Atg8 does not act as a fusogen. Rather, we provide evidence for the involvement of exocytic Q/t-SNAREs in autophagosome formation, acting in the recruitment of key autophagy components to the site of autophagosome formation, and in regulating the organization of Atg9 into tubulovesicular clusters. Additionally, we found that the endosomal Q/t-SNARE Tlg2 and the R/v-SNAREs Sec22 and Ykt6 interact with Sso1-Sec9, and are required for normal Atg9 transport. Thus, multiple SNARE-mediated fusion events are likely to be involved in autophagosome biogenesis.
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