Cargando…
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...
Autores principales: | , , , , , , |
---|---|
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 |
_version_ | 1782311238763544576 |
---|---|
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. |
format | Online Article Text |
id | pubmed-3982997 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2014 |
publisher | The American Society for Cell Biology |
record_format | MEDLINE/PubMed |
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 |
work_keys_str_mv | AT jiangpeidu thehopscomplexmediatesautophagosomelysosomefusionthroughinteractionwithsyntaxin17 AT nishimurataki thehopscomplexmediatesautophagosomelysosomefusionthroughinteractionwithsyntaxin17 AT sakamakiyuriko thehopscomplexmediatesautophagosomelysosomefusionthroughinteractionwithsyntaxin17 AT itakuraeisuke thehopscomplexmediatesautophagosomelysosomefusionthroughinteractionwithsyntaxin17 AT hattatomohisa thehopscomplexmediatesautophagosomelysosomefusionthroughinteractionwithsyntaxin17 AT natsumetohru thehopscomplexmediatesautophagosomelysosomefusionthroughinteractionwithsyntaxin17 AT mizushimanoboru thehopscomplexmediatesautophagosomelysosomefusionthroughinteractionwithsyntaxin17 AT jiangpeidu hopscomplexmediatesautophagosomelysosomefusionthroughinteractionwithsyntaxin17 AT nishimurataki hopscomplexmediatesautophagosomelysosomefusionthroughinteractionwithsyntaxin17 AT sakamakiyuriko hopscomplexmediatesautophagosomelysosomefusionthroughinteractionwithsyntaxin17 AT itakuraeisuke hopscomplexmediatesautophagosomelysosomefusionthroughinteractionwithsyntaxin17 AT hattatomohisa hopscomplexmediatesautophagosomelysosomefusionthroughinteractionwithsyntaxin17 AT natsumetohru hopscomplexmediatesautophagosomelysosomefusionthroughinteractionwithsyntaxin17 AT mizushimanoboru hopscomplexmediatesautophagosomelysosomefusionthroughinteractionwithsyntaxin17 |