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VPS18 recruits VPS41 to the human HOPS complex via a RING–RING interaction
Eukaryotic cells use conserved multisubunit membrane tethering complexes, including CORVET (class C core vacuole/endosome tethering) and HOPS (homotypic fusion and vacuole protein sorting), to control the fusion of endomembranes. These complexes have been extensively studied in yeast, but to date th...
Autores principales: | , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Portland Press Ltd.
2017
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5651818/ https://www.ncbi.nlm.nih.gov/pubmed/28931724 http://dx.doi.org/10.1042/BCJ20170588 |
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author | Hunter, Morag R. Scourfield, Edward J. Emmott, Edward Graham, Stephen C. |
author_facet | Hunter, Morag R. Scourfield, Edward J. Emmott, Edward Graham, Stephen C. |
author_sort | Hunter, Morag R. |
collection | PubMed |
description | Eukaryotic cells use conserved multisubunit membrane tethering complexes, including CORVET (class C core vacuole/endosome tethering) and HOPS (homotypic fusion and vacuole protein sorting), to control the fusion of endomembranes. These complexes have been extensively studied in yeast, but to date there have been far fewer studies of metazoan CORVET and HOPS. Both of these complexes comprise six subunits: a common four-subunit core and two unique subunits. Once assembled, these complexes function to recognise specific endosomal membrane markers and facilitate SNARE-mediated membrane fusion. CORVET promotes the homotypic fusion of early endosomes, while HOPS promotes the fusion of lysosomes to late endosomes and autophagosomes. Many of the subunits of both CORVET and HOPS contain putative C-terminal zinc-finger domains. Here, the contribution of these domains to the assembly of the human CORVET and HOPS complexes has been examined. Using biochemical techniques, we demonstrate that the zinc-containing RING (really interesting new gene) domains of human VPS18 and VPS41 interact directly to form a stable heterodimer. In cells, these RING domains are able to integrate into endogenous HOPS, showing that the VPS18 RING domain is required to recruit VPS41 to the core complex subunits. Importantly, this mechanism is not conserved throughout eukaryotes, as yeast Vps41 does not contain a C-terminal zinc-finger motif. The subunit analogous to VPS41 in human CORVET is VPS8, in which the RING domain has an additional C-terminal segment that is predicted to be disordered. Both the RING and disordered C-terminal domains are required for integration of VPS8 into endogenous CORVET complexes, suggesting that HOPS and CORVET recruit VPS41 and VPS8 via distinct molecular interactions. |
format | Online Article Text |
id | pubmed-5651818 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2017 |
publisher | Portland Press Ltd. |
record_format | MEDLINE/PubMed |
spelling | pubmed-56518182017-10-27 VPS18 recruits VPS41 to the human HOPS complex via a RING–RING interaction Hunter, Morag R. Scourfield, Edward J. Emmott, Edward Graham, Stephen C. Biochem J Research Articles Eukaryotic cells use conserved multisubunit membrane tethering complexes, including CORVET (class C core vacuole/endosome tethering) and HOPS (homotypic fusion and vacuole protein sorting), to control the fusion of endomembranes. These complexes have been extensively studied in yeast, but to date there have been far fewer studies of metazoan CORVET and HOPS. Both of these complexes comprise six subunits: a common four-subunit core and two unique subunits. Once assembled, these complexes function to recognise specific endosomal membrane markers and facilitate SNARE-mediated membrane fusion. CORVET promotes the homotypic fusion of early endosomes, while HOPS promotes the fusion of lysosomes to late endosomes and autophagosomes. Many of the subunits of both CORVET and HOPS contain putative C-terminal zinc-finger domains. Here, the contribution of these domains to the assembly of the human CORVET and HOPS complexes has been examined. Using biochemical techniques, we demonstrate that the zinc-containing RING (really interesting new gene) domains of human VPS18 and VPS41 interact directly to form a stable heterodimer. In cells, these RING domains are able to integrate into endogenous HOPS, showing that the VPS18 RING domain is required to recruit VPS41 to the core complex subunits. Importantly, this mechanism is not conserved throughout eukaryotes, as yeast Vps41 does not contain a C-terminal zinc-finger motif. The subunit analogous to VPS41 in human CORVET is VPS8, in which the RING domain has an additional C-terminal segment that is predicted to be disordered. Both the RING and disordered C-terminal domains are required for integration of VPS8 into endogenous CORVET complexes, suggesting that HOPS and CORVET recruit VPS41 and VPS8 via distinct molecular interactions. Portland Press Ltd. 2017-11-01 2017-10-23 /pmc/articles/PMC5651818/ /pubmed/28931724 http://dx.doi.org/10.1042/BCJ20170588 Text en © 2017 The Author(s) https://creativecommons.org/licenses/by/4.0/This is an open access article published by Portland Press Limited on behalf of the Biochemical Society and distributed under the Creative Commons Attribution License 4.0 (CC BY) (https://creativecommons.org/licenses/by/4.0/) . |
spellingShingle | Research Articles Hunter, Morag R. Scourfield, Edward J. Emmott, Edward Graham, Stephen C. VPS18 recruits VPS41 to the human HOPS complex via a RING–RING interaction |
title | VPS18 recruits VPS41 to the human HOPS complex via a RING–RING interaction |
title_full | VPS18 recruits VPS41 to the human HOPS complex via a RING–RING interaction |
title_fullStr | VPS18 recruits VPS41 to the human HOPS complex via a RING–RING interaction |
title_full_unstemmed | VPS18 recruits VPS41 to the human HOPS complex via a RING–RING interaction |
title_short | VPS18 recruits VPS41 to the human HOPS complex via a RING–RING interaction |
title_sort | vps18 recruits vps41 to the human hops complex via a ring–ring interaction |
topic | Research Articles |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5651818/ https://www.ncbi.nlm.nih.gov/pubmed/28931724 http://dx.doi.org/10.1042/BCJ20170588 |
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