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Regulation of clathrin-mediated endocytosis by hierarchical allosteric activation of AP2

The critical initiation phase of clathrin-mediated endocytosis (CME) determines where and when endocytosis occurs. Heterotetrameric adaptor protein 2 (AP2) complexes, which initiate clathrin-coated pit (CCP) assembly, are activated by conformational changes in response to phosphatidylinositol-4,5-bi...

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Autores principales: Kadlecova, Zuzana, Spielman, Stephanie J., Loerke, Dinah, Mohanakrishnan, Aparna, Reed, Dana Kim, Schmid, Sandra L.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: The Rockefeller University Press 2017
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5223608/
https://www.ncbi.nlm.nih.gov/pubmed/28003333
http://dx.doi.org/10.1083/jcb.201608071
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author Kadlecova, Zuzana
Spielman, Stephanie J.
Loerke, Dinah
Mohanakrishnan, Aparna
Reed, Dana Kim
Schmid, Sandra L.
author_facet Kadlecova, Zuzana
Spielman, Stephanie J.
Loerke, Dinah
Mohanakrishnan, Aparna
Reed, Dana Kim
Schmid, Sandra L.
author_sort Kadlecova, Zuzana
collection PubMed
description The critical initiation phase of clathrin-mediated endocytosis (CME) determines where and when endocytosis occurs. Heterotetrameric adaptor protein 2 (AP2) complexes, which initiate clathrin-coated pit (CCP) assembly, are activated by conformational changes in response to phosphatidylinositol-4,5-bisphosphate (PIP2) and cargo binding at multiple sites. However, the functional hierarchy of interactions and how these conformational changes relate to distinct steps in CCP formation in living cells remains unknown. We used quantitative live-cell analyses to measure discrete early stages of CME and show how sequential, allosterically regulated conformational changes activate AP2 to drive both nucleation and subsequent stabilization of nascent CCPs. Our data establish that cargoes containing Yxxφ motif, but not dileucine motif, play a critical role in the earliest stages of AP2 activation and CCP nucleation. Interestingly, these cargo and PIP2 interactions are not conserved in yeast. Thus, we speculate that AP2 has evolved as a key regulatory node to coordinate CCP formation and cargo sorting and ensure high spatial and temporal regulation of CME.
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spelling pubmed-52236082017-07-02 Regulation of clathrin-mediated endocytosis by hierarchical allosteric activation of AP2 Kadlecova, Zuzana Spielman, Stephanie J. Loerke, Dinah Mohanakrishnan, Aparna Reed, Dana Kim Schmid, Sandra L. J Cell Biol Research Articles The critical initiation phase of clathrin-mediated endocytosis (CME) determines where and when endocytosis occurs. Heterotetrameric adaptor protein 2 (AP2) complexes, which initiate clathrin-coated pit (CCP) assembly, are activated by conformational changes in response to phosphatidylinositol-4,5-bisphosphate (PIP2) and cargo binding at multiple sites. However, the functional hierarchy of interactions and how these conformational changes relate to distinct steps in CCP formation in living cells remains unknown. We used quantitative live-cell analyses to measure discrete early stages of CME and show how sequential, allosterically regulated conformational changes activate AP2 to drive both nucleation and subsequent stabilization of nascent CCPs. Our data establish that cargoes containing Yxxφ motif, but not dileucine motif, play a critical role in the earliest stages of AP2 activation and CCP nucleation. Interestingly, these cargo and PIP2 interactions are not conserved in yeast. Thus, we speculate that AP2 has evolved as a key regulatory node to coordinate CCP formation and cargo sorting and ensure high spatial and temporal regulation of CME. The Rockefeller University Press 2017-01-02 /pmc/articles/PMC5223608/ /pubmed/28003333 http://dx.doi.org/10.1083/jcb.201608071 Text en © 2017 Kadlecova et al. https://creativecommons.org/licenses/by-nc-sa/4.0/http://www.rupress.org/terms/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
Kadlecova, Zuzana
Spielman, Stephanie J.
Loerke, Dinah
Mohanakrishnan, Aparna
Reed, Dana Kim
Schmid, Sandra L.
Regulation of clathrin-mediated endocytosis by hierarchical allosteric activation of AP2
title Regulation of clathrin-mediated endocytosis by hierarchical allosteric activation of AP2
title_full Regulation of clathrin-mediated endocytosis by hierarchical allosteric activation of AP2
title_fullStr Regulation of clathrin-mediated endocytosis by hierarchical allosteric activation of AP2
title_full_unstemmed Regulation of clathrin-mediated endocytosis by hierarchical allosteric activation of AP2
title_short Regulation of clathrin-mediated endocytosis by hierarchical allosteric activation of AP2
title_sort regulation of clathrin-mediated endocytosis by hierarchical allosteric activation of ap2
topic Research Articles
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5223608/
https://www.ncbi.nlm.nih.gov/pubmed/28003333
http://dx.doi.org/10.1083/jcb.201608071
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