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Iterative sorting of apical and basolateral cargo in Madin–Darby canine kidney cells
For several decades, the trans-Golgi network (TGN) was considered the most distal stop and hence the ultimate protein-sorting station for distinct apical and basolateral transport carriers that reach their respective surface domains in the direct trafficking pathway. However, recent reports of apica...
Autores principales: | , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
The American Society for Cell Biology
2016
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4945143/ https://www.ncbi.nlm.nih.gov/pubmed/27226480 http://dx.doi.org/10.1091/mbc.E16-02-0096 |
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author | Treyer, Aleksandr Pujato, Mario Pechuan, Ximo Müsch, Anne |
author_facet | Treyer, Aleksandr Pujato, Mario Pechuan, Ximo Müsch, Anne |
author_sort | Treyer, Aleksandr |
collection | PubMed |
description | For several decades, the trans-Golgi network (TGN) was considered the most distal stop and hence the ultimate protein-sorting station for distinct apical and basolateral transport carriers that reach their respective surface domains in the direct trafficking pathway. However, recent reports of apical and basolateral cargoes traversing post-Golgi compartments accessible to endocytic ligands before their arrival at the cell surface and the post-TGN breakup of large pleomorphic membrane fragments that exit the Golgi region toward the surface raised the possibility that compartments distal to the TGN mediate or contribute to biosynthetic sorting. Here we describe the development of a novel assay that quantitatively distinguishes different cargo pairs by their degree of colocalization at the TGN and by the evolution of colocalization during their TGN-to-surface transport. Keys to the high resolution of our approach are 1) conversion of perinuclear organelle clustering into a two-dimensional microsomal spread and 2) identification of TGN and post-TGN cargo without the need for a TGN marker that universally cosegregates with all cargo. Using our assay, we provide the first evidence that apical NTRp75 and basolateral VSVG in Madin–Darby canine kidney cells still undergo progressive sorting after they exit the TGN toward the cell surface. |
format | Online Article Text |
id | pubmed-4945143 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2016 |
publisher | The American Society for Cell Biology |
record_format | MEDLINE/PubMed |
spelling | pubmed-49451432016-09-30 Iterative sorting of apical and basolateral cargo in Madin–Darby canine kidney cells Treyer, Aleksandr Pujato, Mario Pechuan, Ximo Müsch, Anne Mol Biol Cell Articles For several decades, the trans-Golgi network (TGN) was considered the most distal stop and hence the ultimate protein-sorting station for distinct apical and basolateral transport carriers that reach their respective surface domains in the direct trafficking pathway. However, recent reports of apical and basolateral cargoes traversing post-Golgi compartments accessible to endocytic ligands before their arrival at the cell surface and the post-TGN breakup of large pleomorphic membrane fragments that exit the Golgi region toward the surface raised the possibility that compartments distal to the TGN mediate or contribute to biosynthetic sorting. Here we describe the development of a novel assay that quantitatively distinguishes different cargo pairs by their degree of colocalization at the TGN and by the evolution of colocalization during their TGN-to-surface transport. Keys to the high resolution of our approach are 1) conversion of perinuclear organelle clustering into a two-dimensional microsomal spread and 2) identification of TGN and post-TGN cargo without the need for a TGN marker that universally cosegregates with all cargo. Using our assay, we provide the first evidence that apical NTRp75 and basolateral VSVG in Madin–Darby canine kidney cells still undergo progressive sorting after they exit the TGN toward the cell surface. The American Society for Cell Biology 2016-07-15 /pmc/articles/PMC4945143/ /pubmed/27226480 http://dx.doi.org/10.1091/mbc.E16-02-0096 Text en © 2016 Treyer 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 for Cell Biology. |
spellingShingle | Articles Treyer, Aleksandr Pujato, Mario Pechuan, Ximo Müsch, Anne Iterative sorting of apical and basolateral cargo in Madin–Darby canine kidney cells |
title | Iterative sorting of apical and basolateral cargo in Madin–Darby canine kidney cells |
title_full | Iterative sorting of apical and basolateral cargo in Madin–Darby canine kidney cells |
title_fullStr | Iterative sorting of apical and basolateral cargo in Madin–Darby canine kidney cells |
title_full_unstemmed | Iterative sorting of apical and basolateral cargo in Madin–Darby canine kidney cells |
title_short | Iterative sorting of apical and basolateral cargo in Madin–Darby canine kidney cells |
title_sort | iterative sorting of apical and basolateral cargo in madin–darby canine kidney cells |
topic | Articles |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4945143/ https://www.ncbi.nlm.nih.gov/pubmed/27226480 http://dx.doi.org/10.1091/mbc.E16-02-0096 |
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