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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...

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Detalles Bibliográficos
Autores principales: Nelms, Bradlee, Dalomba, Natasha Furtado, Lencer, Wayne
Formato: Online Artículo Texto
Lenguaje:English
Publicado: The Rockefeller University Press 2017
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.
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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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