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The HOPS complex mediates autophagosome–lysosome fusion through interaction with syntaxin 17

Membrane fusion is generally controlled by Rabs, soluble N-ethylmaleimide–sensitive factor attachment protein receptors (SNAREs), and tethering complexes. Syntaxin 17 (STX17) was recently identified as the autophagosomal SNARE required for autophagosome–lysosome fusion in mammals and Drosophila. In...

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Autores principales: Jiang, Peidu, Nishimura, Taki, Sakamaki, Yuriko, Itakura, Eisuke, Hatta, Tomohisa, Natsume, Tohru, Mizushima, Noboru
Formato: Online Artículo Texto
Lenguaje:English
Publicado: The American Society for Cell Biology 2014
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3982997/
https://www.ncbi.nlm.nih.gov/pubmed/24554770
http://dx.doi.org/10.1091/mbc.E13-08-0447
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author Jiang, Peidu
Nishimura, Taki
Sakamaki, Yuriko
Itakura, Eisuke
Hatta, Tomohisa
Natsume, Tohru
Mizushima, Noboru
author_facet Jiang, Peidu
Nishimura, Taki
Sakamaki, Yuriko
Itakura, Eisuke
Hatta, Tomohisa
Natsume, Tohru
Mizushima, Noboru
author_sort Jiang, Peidu
collection PubMed
description Membrane fusion is generally controlled by Rabs, soluble N-ethylmaleimide–sensitive factor attachment protein receptors (SNAREs), and tethering complexes. Syntaxin 17 (STX17) was recently identified as the autophagosomal SNARE required for autophagosome–lysosome fusion in mammals and Drosophila. In this study, to better understand the mechanism of autophagosome–lysosome fusion, we searched for STX17-interacting proteins. Immunoprecipitation and mass spectrometry analysis identified vacuolar protein sorting 33A (VPS33A) and VPS16, which are components of the homotypic fusion and protein sorting (HOPS)–tethering complex. We further confirmed that all HOPS components were coprecipitated with STX17. Knockdown of VPS33A, VPS16, or VPS39 blocked autophagic flux and caused accumulation of STX17- and microtubule-associated protein light chain (LC3)–positive autophagosomes. The endocytic pathway was also affected by knockdown of VPS33A, as previously reported, but not by knockdown of STX17. By contrast, ultraviolet irradiation resistance–associated gene (UVRAG), a known HOPS-interacting protein, did not interact with the STX17–HOPS complex and may not be directly involved in autophagosome–lysosome fusion. Collectively these results suggest that, in addition to its well-established function in the endocytic pathway, HOPS promotes autophagosome–lysosome fusion through interaction with STX17.
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spelling pubmed-39829972014-06-30 The HOPS complex mediates autophagosome–lysosome fusion through interaction with syntaxin 17 Jiang, Peidu Nishimura, Taki Sakamaki, Yuriko Itakura, Eisuke Hatta, Tomohisa Natsume, Tohru Mizushima, Noboru Mol Biol Cell Articles Membrane fusion is generally controlled by Rabs, soluble N-ethylmaleimide–sensitive factor attachment protein receptors (SNAREs), and tethering complexes. Syntaxin 17 (STX17) was recently identified as the autophagosomal SNARE required for autophagosome–lysosome fusion in mammals and Drosophila. In this study, to better understand the mechanism of autophagosome–lysosome fusion, we searched for STX17-interacting proteins. Immunoprecipitation and mass spectrometry analysis identified vacuolar protein sorting 33A (VPS33A) and VPS16, which are components of the homotypic fusion and protein sorting (HOPS)–tethering complex. We further confirmed that all HOPS components were coprecipitated with STX17. Knockdown of VPS33A, VPS16, or VPS39 blocked autophagic flux and caused accumulation of STX17- and microtubule-associated protein light chain (LC3)–positive autophagosomes. The endocytic pathway was also affected by knockdown of VPS33A, as previously reported, but not by knockdown of STX17. By contrast, ultraviolet irradiation resistance–associated gene (UVRAG), a known HOPS-interacting protein, did not interact with the STX17–HOPS complex and may not be directly involved in autophagosome–lysosome fusion. Collectively these results suggest that, in addition to its well-established function in the endocytic pathway, HOPS promotes autophagosome–lysosome fusion through interaction with STX17. The American Society for Cell Biology 2014-04-15 /pmc/articles/PMC3982997/ /pubmed/24554770 http://dx.doi.org/10.1091/mbc.E13-08-0447 Text en © 2014 Jiang et al. This article is distributed by The American Society for Cell Biology under license from the author(s). Two months after publication it is available to the public under an Attribution–Noncommercial–Share Alike 3.0 Unported Creative Commons License (http://creativecommons.org/licenses/by-nc-sa/3.0). “ASCB®,” “The American Society for Cell Biology®,” and “Molecular Biology of the Cell®” are registered trademarks of The American Society of Cell Biology.
spellingShingle Articles
Jiang, Peidu
Nishimura, Taki
Sakamaki, Yuriko
Itakura, Eisuke
Hatta, Tomohisa
Natsume, Tohru
Mizushima, Noboru
The HOPS complex mediates autophagosome–lysosome fusion through interaction with syntaxin 17
title The HOPS complex mediates autophagosome–lysosome fusion through interaction with syntaxin 17
title_full The HOPS complex mediates autophagosome–lysosome fusion through interaction with syntaxin 17
title_fullStr The HOPS complex mediates autophagosome–lysosome fusion through interaction with syntaxin 17
title_full_unstemmed The HOPS complex mediates autophagosome–lysosome fusion through interaction with syntaxin 17
title_short The HOPS complex mediates autophagosome–lysosome fusion through interaction with syntaxin 17
title_sort hops complex mediates autophagosome–lysosome fusion through interaction with syntaxin 17
topic Articles
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3982997/
https://www.ncbi.nlm.nih.gov/pubmed/24554770
http://dx.doi.org/10.1091/mbc.E13-08-0447
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