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A microscopy-based kinetic analysis of yeast vacuolar protein sorting
Saccharomyces cerevisiae is amenable to studying membrane traffic by live-cell fluorescence microscopy. We used this system to explore two aspects of cargo protein traffic through prevacuolar endosome (PVE) compartments to the vacuole. First, at what point during Golgi maturation does a biosynthetic...
Autores principales: | , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
eLife Sciences Publications, Ltd
2020
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7338053/ https://www.ncbi.nlm.nih.gov/pubmed/32584255 http://dx.doi.org/10.7554/eLife.56844 |
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author | Casler, Jason C Glick, Benjamin S |
author_facet | Casler, Jason C Glick, Benjamin S |
author_sort | Casler, Jason C |
collection | PubMed |
description | Saccharomyces cerevisiae is amenable to studying membrane traffic by live-cell fluorescence microscopy. We used this system to explore two aspects of cargo protein traffic through prevacuolar endosome (PVE) compartments to the vacuole. First, at what point during Golgi maturation does a biosynthetic vacuolar cargo depart from the maturing cisternae? To address this question, we modified a regulatable fluorescent secretory cargo by adding a vacuolar targeting signal. Traffic of the vacuolar cargo requires the GGA clathrin adaptors, which arrive during the early-to-late Golgi transition. Accordingly, the vacuolar cargo begins to exit the Golgi near the midpoint of maturation, significantly before exit of a secretory cargo. Second, how are cargoes delivered from PVE compartments to the vacuole? To address this question, we tracked biosynthetic and endocytic cargoes after they had accumulated in PVE compartments. The results suggest that stable PVE compartments repeatedly deliver material to the vacuole by a kiss-and-run mechanism. |
format | Online Article Text |
id | pubmed-7338053 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2020 |
publisher | eLife Sciences Publications, Ltd |
record_format | MEDLINE/PubMed |
spelling | pubmed-73380532020-07-13 A microscopy-based kinetic analysis of yeast vacuolar protein sorting Casler, Jason C Glick, Benjamin S eLife Cell Biology Saccharomyces cerevisiae is amenable to studying membrane traffic by live-cell fluorescence microscopy. We used this system to explore two aspects of cargo protein traffic through prevacuolar endosome (PVE) compartments to the vacuole. First, at what point during Golgi maturation does a biosynthetic vacuolar cargo depart from the maturing cisternae? To address this question, we modified a regulatable fluorescent secretory cargo by adding a vacuolar targeting signal. Traffic of the vacuolar cargo requires the GGA clathrin adaptors, which arrive during the early-to-late Golgi transition. Accordingly, the vacuolar cargo begins to exit the Golgi near the midpoint of maturation, significantly before exit of a secretory cargo. Second, how are cargoes delivered from PVE compartments to the vacuole? To address this question, we tracked biosynthetic and endocytic cargoes after they had accumulated in PVE compartments. The results suggest that stable PVE compartments repeatedly deliver material to the vacuole by a kiss-and-run mechanism. eLife Sciences Publications, Ltd 2020-06-25 /pmc/articles/PMC7338053/ /pubmed/32584255 http://dx.doi.org/10.7554/eLife.56844 Text en © 2020, Casler and Glick https://creativecommons.org/licenses/by/4.0/This article is distributed under the terms of the Creative Commons Attribution License (https://creativecommons.org/licenses/by/4.0/) , which permits unrestricted use and redistribution provided that the original author and source are credited. |
spellingShingle | Cell Biology Casler, Jason C Glick, Benjamin S A microscopy-based kinetic analysis of yeast vacuolar protein sorting |
title | A microscopy-based kinetic analysis of yeast vacuolar protein sorting |
title_full | A microscopy-based kinetic analysis of yeast vacuolar protein sorting |
title_fullStr | A microscopy-based kinetic analysis of yeast vacuolar protein sorting |
title_full_unstemmed | A microscopy-based kinetic analysis of yeast vacuolar protein sorting |
title_short | A microscopy-based kinetic analysis of yeast vacuolar protein sorting |
title_sort | microscopy-based kinetic analysis of yeast vacuolar protein sorting |
topic | Cell Biology |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7338053/ https://www.ncbi.nlm.nih.gov/pubmed/32584255 http://dx.doi.org/10.7554/eLife.56844 |
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