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New tools for “hot-wiring” clathrin-mediated endocytosis with temporal and spatial precision
Clathrin-mediated endocytosis (CME) is the major route of receptor internalization at the plasma membrane. Analysis of constitutive CME is difficult because the initiation of endocytic events is unpredictable. When and where a clathrin-coated pit will form and what cargo it will contain are difficul...
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/PMC5716275/ https://www.ncbi.nlm.nih.gov/pubmed/28954824 http://dx.doi.org/10.1083/jcb.201702188 |
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author | Wood, Laura A. Larocque, Gabrielle Clarke, Nicholas I. Sarkar, Sourav Royle, Stephen J. |
author_facet | Wood, Laura A. Larocque, Gabrielle Clarke, Nicholas I. Sarkar, Sourav Royle, Stephen J. |
author_sort | Wood, Laura A. |
collection | PubMed |
description | Clathrin-mediated endocytosis (CME) is the major route of receptor internalization at the plasma membrane. Analysis of constitutive CME is difficult because the initiation of endocytic events is unpredictable. When and where a clathrin-coated pit will form and what cargo it will contain are difficult to foresee. Here we describe a series of genetically encoded reporters that allow the initiation of CME on demand. A clathrin-binding protein fragment (“hook”) is inducibly attached to an “anchor” protein at the plasma membrane, which triggers the formation of new clathrin-coated vesicles. Our design incorporates temporal and spatial control by the use of chemical and optogenetic methods for inducing hook–anchor attachment. Moreover, the cargo is defined. Because several steps in vesicle creation are bypassed, we term it “hot-wiring.” We use hot-wired endocytosis to describe the functional interactions between clathrin and AP2. Two distinct sites on the β2 subunit, one on the hinge and the other on the appendage, are necessary and sufficient for functional clathrin engagement. |
format | Online Article Text |
id | pubmed-5716275 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2017 |
publisher | The Rockefeller University Press |
record_format | MEDLINE/PubMed |
spelling | pubmed-57162752018-06-04 New tools for “hot-wiring” clathrin-mediated endocytosis with temporal and spatial precision Wood, Laura A. Larocque, Gabrielle Clarke, Nicholas I. Sarkar, Sourav Royle, Stephen J. J Cell Biol Research Articles Clathrin-mediated endocytosis (CME) is the major route of receptor internalization at the plasma membrane. Analysis of constitutive CME is difficult because the initiation of endocytic events is unpredictable. When and where a clathrin-coated pit will form and what cargo it will contain are difficult to foresee. Here we describe a series of genetically encoded reporters that allow the initiation of CME on demand. A clathrin-binding protein fragment (“hook”) is inducibly attached to an “anchor” protein at the plasma membrane, which triggers the formation of new clathrin-coated vesicles. Our design incorporates temporal and spatial control by the use of chemical and optogenetic methods for inducing hook–anchor attachment. Moreover, the cargo is defined. Because several steps in vesicle creation are bypassed, we term it “hot-wiring.” We use hot-wired endocytosis to describe the functional interactions between clathrin and AP2. Two distinct sites on the β2 subunit, one on the hinge and the other on the appendage, are necessary and sufficient for functional clathrin engagement. The Rockefeller University Press 2017-12-04 /pmc/articles/PMC5716275/ /pubmed/28954824 http://dx.doi.org/10.1083/jcb.201702188 Text en © 2017 Wood et al. http://www.rupress.org/terms/https://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/). 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 Wood, Laura A. Larocque, Gabrielle Clarke, Nicholas I. Sarkar, Sourav Royle, Stephen J. New tools for “hot-wiring” clathrin-mediated endocytosis with temporal and spatial precision |
title | New tools for “hot-wiring” clathrin-mediated endocytosis with temporal and spatial precision |
title_full | New tools for “hot-wiring” clathrin-mediated endocytosis with temporal and spatial precision |
title_fullStr | New tools for “hot-wiring” clathrin-mediated endocytosis with temporal and spatial precision |
title_full_unstemmed | New tools for “hot-wiring” clathrin-mediated endocytosis with temporal and spatial precision |
title_short | New tools for “hot-wiring” clathrin-mediated endocytosis with temporal and spatial precision |
title_sort | new tools for “hot-wiring” clathrin-mediated endocytosis with temporal and spatial precision |
topic | Research Articles |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5716275/ https://www.ncbi.nlm.nih.gov/pubmed/28954824 http://dx.doi.org/10.1083/jcb.201702188 |
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