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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...

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Detalles Bibliográficos
Autores principales: Gao, Jieqiong, Reggiori, Fulvio, Ungermann, Christian
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Rockefeller University Press 2018
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.
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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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