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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...

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Detalles Bibliográficos
Autores principales: Casler, Jason C, Glick, Benjamin S
Formato: Online Artículo Texto
Lenguaje:English
Publicado: eLife Sciences Publications, Ltd 2020
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.
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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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