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Ultrastructure of COPII vesicle formation in yeast characterized by correlative light and electron microscopy

Traffic of proteins out of the endoplasmic reticulum (ER) is driven by the COPII coat, a layered protein scaffold that mediates the capture of cargo proteins and the remodeling of the ER membrane into spherical vesicular carriers. Although the components of this machinery have been genetically defin...

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Autores principales: Melero, Alejandro, Boulanger, Jerome, Kukulski, Wanda, Miller, Elizabeth A.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: The American Society for Cell Biology 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9634970/
https://www.ncbi.nlm.nih.gov/pubmed/36001360
http://dx.doi.org/10.1091/mbc.E22-03-0103
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author Melero, Alejandro
Boulanger, Jerome
Kukulski, Wanda
Miller, Elizabeth A.
author_facet Melero, Alejandro
Boulanger, Jerome
Kukulski, Wanda
Miller, Elizabeth A.
author_sort Melero, Alejandro
collection PubMed
description Traffic of proteins out of the endoplasmic reticulum (ER) is driven by the COPII coat, a layered protein scaffold that mediates the capture of cargo proteins and the remodeling of the ER membrane into spherical vesicular carriers. Although the components of this machinery have been genetically defined, and the mechanisms of coat assembly extensively explored in vitro, understanding the physical mechanisms of membrane remodeling in cells remains a challenge. Here we use correlative light and electron microscopy (CLEM) to visualize the nanoscale ultrastructure of membrane remodeling at ER exit sites (ERES) in yeast cells. Using various COPII mutants, we have determined the broad contribution that each layer of the coat makes to membrane remodeling. Our data suggest that inner coat components define the radius of curvature, whereas outer coat components facilitate membrane fission. The organization of the coat in conjunction with membrane biophysical properties determines the ultrastructure of vesicles and thus the efficiency of protein transport.
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spelling pubmed-96349702023-01-16 Ultrastructure of COPII vesicle formation in yeast characterized by correlative light and electron microscopy Melero, Alejandro Boulanger, Jerome Kukulski, Wanda Miller, Elizabeth A. Mol Biol Cell Articles Traffic of proteins out of the endoplasmic reticulum (ER) is driven by the COPII coat, a layered protein scaffold that mediates the capture of cargo proteins and the remodeling of the ER membrane into spherical vesicular carriers. Although the components of this machinery have been genetically defined, and the mechanisms of coat assembly extensively explored in vitro, understanding the physical mechanisms of membrane remodeling in cells remains a challenge. Here we use correlative light and electron microscopy (CLEM) to visualize the nanoscale ultrastructure of membrane remodeling at ER exit sites (ERES) in yeast cells. Using various COPII mutants, we have determined the broad contribution that each layer of the coat makes to membrane remodeling. Our data suggest that inner coat components define the radius of curvature, whereas outer coat components facilitate membrane fission. The organization of the coat in conjunction with membrane biophysical properties determines the ultrastructure of vesicles and thus the efficiency of protein transport. The American Society for Cell Biology 2022-11-01 /pmc/articles/PMC9634970/ /pubmed/36001360 http://dx.doi.org/10.1091/mbc.E22-03-0103 Text en © 2022 Melero et al. “ASCB®,” “The American Society for Cell Biology®,” and “Molecular Biology of the Cell®” are registered trademarks of The American Society for Cell Biology. https://creativecommons.org/licenses/by-nc-sa/4.0/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 4.0 International Creative Commons License.
spellingShingle Articles
Melero, Alejandro
Boulanger, Jerome
Kukulski, Wanda
Miller, Elizabeth A.
Ultrastructure of COPII vesicle formation in yeast characterized by correlative light and electron microscopy
title Ultrastructure of COPII vesicle formation in yeast characterized by correlative light and electron microscopy
title_full Ultrastructure of COPII vesicle formation in yeast characterized by correlative light and electron microscopy
title_fullStr Ultrastructure of COPII vesicle formation in yeast characterized by correlative light and electron microscopy
title_full_unstemmed Ultrastructure of COPII vesicle formation in yeast characterized by correlative light and electron microscopy
title_short Ultrastructure of COPII vesicle formation in yeast characterized by correlative light and electron microscopy
title_sort ultrastructure of copii vesicle formation in yeast characterized by correlative light and electron microscopy
topic Articles
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9634970/
https://www.ncbi.nlm.nih.gov/pubmed/36001360
http://dx.doi.org/10.1091/mbc.E22-03-0103
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