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A novel in vitro assay reveals SNARE topology and the role of Ykt6 in autophagosome fusion with vacuoles
Autophagy is a catabolic pathway that delivers intracellular material to the mammalian lysosomes or the yeast and plant vacuoles. The final step in this process is the fusion of autophagosomes with vacuoles, which requires SNARE proteins, the homotypic vacuole fusion and protein sorting tethering co...
Autores principales: | , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Rockefeller University Press
2018
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6168247/ https://www.ncbi.nlm.nih.gov/pubmed/30097515 http://dx.doi.org/10.1083/jcb.201804039 |
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author | Gao, Jieqiong Reggiori, Fulvio Ungermann, Christian |
author_facet | Gao, Jieqiong Reggiori, Fulvio Ungermann, Christian |
author_sort | Gao, Jieqiong |
collection | PubMed |
description | Autophagy is a catabolic pathway that delivers intracellular material to the mammalian lysosomes or the yeast and plant vacuoles. The final step in this process is the fusion of autophagosomes with vacuoles, which requires SNARE proteins, the homotypic vacuole fusion and protein sorting tethering complex, the RAB7-like Ypt7 GTPase, and its guanine nucleotide exchange factor, Mon1-Ccz1. Where these different components are located and function during fusion, however, remains to be fully understood. Here, we present a novel in vitro assay to monitor fusion of intact and functional autophagosomes with vacuoles. This process requires ATP, physiological temperature, and the entire fusion machinery to tether and fuse autophagosomes with vacuoles. Importantly, we uncover Ykt6 as the autophagosomal SNARE. Our assay and findings thus provide the tools to dissect autophagosome completion and fusion in a test tube. |
format | Online Article Text |
id | pubmed-6168247 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2018 |
publisher | Rockefeller University Press |
record_format | MEDLINE/PubMed |
spelling | pubmed-61682472019-04-01 A novel in vitro assay reveals SNARE topology and the role of Ykt6 in autophagosome fusion with vacuoles Gao, Jieqiong Reggiori, Fulvio Ungermann, Christian J Cell Biol Research Articles Autophagy is a catabolic pathway that delivers intracellular material to the mammalian lysosomes or the yeast and plant vacuoles. The final step in this process is the fusion of autophagosomes with vacuoles, which requires SNARE proteins, the homotypic vacuole fusion and protein sorting tethering complex, the RAB7-like Ypt7 GTPase, and its guanine nucleotide exchange factor, Mon1-Ccz1. Where these different components are located and function during fusion, however, remains to be fully understood. Here, we present a novel in vitro assay to monitor fusion of intact and functional autophagosomes with vacuoles. This process requires ATP, physiological temperature, and the entire fusion machinery to tether and fuse autophagosomes with vacuoles. Importantly, we uncover Ykt6 as the autophagosomal SNARE. Our assay and findings thus provide the tools to dissect autophagosome completion and fusion in a test tube. Rockefeller University Press 2018-10-01 /pmc/articles/PMC6168247/ /pubmed/30097515 http://dx.doi.org/10.1083/jcb.201804039 Text en © 2018 Gao et al. http://www.rupress.org/terms/https://creativecommons.org/licenses/by-nc-sa/4.0/This article is distributed under the terms of an Attribution–Noncommercial–Share Alike–No Mirror Sites license for the first six months after the publication date (see http://www.rupress.org/terms/). After six months it is available under a Creative Commons License (Attribution–Noncommercial–Share Alike 4.0 International license, as described at https://creativecommons.org/licenses/by-nc-sa/4.0/). |
spellingShingle | Research Articles Gao, Jieqiong Reggiori, Fulvio Ungermann, Christian A novel in vitro assay reveals SNARE topology and the role of Ykt6 in autophagosome fusion with vacuoles |
title | A novel in vitro assay reveals SNARE topology and the role of Ykt6 in autophagosome fusion with vacuoles |
title_full | A novel in vitro assay reveals SNARE topology and the role of Ykt6 in autophagosome fusion with vacuoles |
title_fullStr | A novel in vitro assay reveals SNARE topology and the role of Ykt6 in autophagosome fusion with vacuoles |
title_full_unstemmed | A novel in vitro assay reveals SNARE topology and the role of Ykt6 in autophagosome fusion with vacuoles |
title_short | A novel in vitro assay reveals SNARE topology and the role of Ykt6 in autophagosome fusion with vacuoles |
title_sort | novel in vitro assay reveals snare topology and the role of ykt6 in autophagosome fusion with vacuoles |
topic | Research Articles |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6168247/ https://www.ncbi.nlm.nih.gov/pubmed/30097515 http://dx.doi.org/10.1083/jcb.201804039 |
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