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A targeted RNAi screen identifies factors affecting diverse stages of receptor-mediated transcytosis
Endosome transport by transcytosis is the primary mechanism by which proteins and other large cargo traverse epithelial barriers in normal tissue. Transcytosis is also essential for establishing and maintaining membrane polarity in epithelia and other polarized cells. To identify novel components of...
Autores principales: | , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
The Rockefeller University Press
2017
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5294788/ https://www.ncbi.nlm.nih.gov/pubmed/28069747 http://dx.doi.org/10.1083/jcb.201609035 |
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author | Nelms, Bradlee Dalomba, Natasha Furtado Lencer, Wayne |
author_facet | Nelms, Bradlee Dalomba, Natasha Furtado Lencer, Wayne |
author_sort | Nelms, Bradlee |
collection | PubMed |
description | Endosome transport by transcytosis is the primary mechanism by which proteins and other large cargo traverse epithelial barriers in normal tissue. Transcytosis is also essential for establishing and maintaining membrane polarity in epithelia and other polarized cells. To identify novel components of this pathway, we conducted a high-throughput RNA interference screen for factors necessary for the bidirectional transcytosis of IgG by the Fcγ receptor FcRn. This screen identified 23 genes whose suppression resulted in a reproducible decrease in FcRn-mediated transcytosis. Pulse-chase kinetic transport assays on four of the top-ranking genes (EXOC2, EXOC7, PARD6B, and LEPROT) revealed distinct effects on the apical and basolateral recycling and transcytotic pathways, demonstrating that these pathways are genetically separable. We also found a strong dependence on PARD6B for apical, but not basolateral, recycling, implicating this cell polarity gene in assembly or maintenance of the apical endosomal system. This dataset yields insights into how vesicular transport is adapted to the specialized functions of differentiated cell types and opens new research avenues into epithelial trafficking. |
format | Online Article Text |
id | pubmed-5294788 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2017 |
publisher | The Rockefeller University Press |
record_format | MEDLINE/PubMed |
spelling | pubmed-52947882017-08-01 A targeted RNAi screen identifies factors affecting diverse stages of receptor-mediated transcytosis Nelms, Bradlee Dalomba, Natasha Furtado Lencer, Wayne J Cell Biol Research Articles Endosome transport by transcytosis is the primary mechanism by which proteins and other large cargo traverse epithelial barriers in normal tissue. Transcytosis is also essential for establishing and maintaining membrane polarity in epithelia and other polarized cells. To identify novel components of this pathway, we conducted a high-throughput RNA interference screen for factors necessary for the bidirectional transcytosis of IgG by the Fcγ receptor FcRn. This screen identified 23 genes whose suppression resulted in a reproducible decrease in FcRn-mediated transcytosis. Pulse-chase kinetic transport assays on four of the top-ranking genes (EXOC2, EXOC7, PARD6B, and LEPROT) revealed distinct effects on the apical and basolateral recycling and transcytotic pathways, demonstrating that these pathways are genetically separable. We also found a strong dependence on PARD6B for apical, but not basolateral, recycling, implicating this cell polarity gene in assembly or maintenance of the apical endosomal system. This dataset yields insights into how vesicular transport is adapted to the specialized functions of differentiated cell types and opens new research avenues into epithelial trafficking. The Rockefeller University Press 2017-02 /pmc/articles/PMC5294788/ /pubmed/28069747 http://dx.doi.org/10.1083/jcb.201609035 Text en © 2017 Nelms et al. http://www.rupress.org/termshttps://creativecommons.org/licenses/by-nc-sa/4.0/This article is distributed under the terms of an Attribution–Noncommercial–Share Alike–No Mirror Sites license for the first six months after the publication date (see http://www.rupress.org/terms (http://www.rupress.org/terms/) ). After six months it is available under a Creative Commons License (Attribution–Noncommercial–Share Alike 4.0 International license, as described at https://creativecommons.org/licenses/by-nc-sa/4.0/). |
spellingShingle | Research Articles Nelms, Bradlee Dalomba, Natasha Furtado Lencer, Wayne A targeted RNAi screen identifies factors affecting diverse stages of receptor-mediated transcytosis |
title | A targeted RNAi screen identifies factors affecting diverse stages of receptor-mediated transcytosis |
title_full | A targeted RNAi screen identifies factors affecting diverse stages of receptor-mediated transcytosis |
title_fullStr | A targeted RNAi screen identifies factors affecting diverse stages of receptor-mediated transcytosis |
title_full_unstemmed | A targeted RNAi screen identifies factors affecting diverse stages of receptor-mediated transcytosis |
title_short | A targeted RNAi screen identifies factors affecting diverse stages of receptor-mediated transcytosis |
title_sort | targeted rnai screen identifies factors affecting diverse stages of receptor-mediated transcytosis |
topic | Research Articles |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5294788/ https://www.ncbi.nlm.nih.gov/pubmed/28069747 http://dx.doi.org/10.1083/jcb.201609035 |
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