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An endosomal tether undergoes an entropic collapse to bring vesicles together

An early step in intracellular transport is the selective recognition of a vesicle by its appropriate target membrane, a process regulated by Rab GTPases via the recruitment of tethering effectors1–4. Membrane tethering confers higher selectivity and efficiency to membrane fusion than the pairing of...

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Autores principales: Murray, David H., Jahnel, Marcus, Lauer, Janelle, Avellaneda, Mario J., Brouilly, Nicolas, Cezanne, Alice, Morales-Navarrete, Hernán, Perini, Enrico D., Ferguson, Charles, Lupas, Andrei N., Kalaidzidis, Yannis, Parton, Robert G., Grill, Stephan W., Zerial, Marino
Formato: Online Artículo Texto
Lenguaje:English
Publicado: 2016
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5142606/
https://www.ncbi.nlm.nih.gov/pubmed/27556945
http://dx.doi.org/10.1038/nature19326
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author Murray, David H.
Jahnel, Marcus
Lauer, Janelle
Avellaneda, Mario J.
Brouilly, Nicolas
Cezanne, Alice
Morales-Navarrete, Hernán
Perini, Enrico D.
Ferguson, Charles
Lupas, Andrei N.
Kalaidzidis, Yannis
Parton, Robert G.
Grill, Stephan W.
Zerial, Marino
author_facet Murray, David H.
Jahnel, Marcus
Lauer, Janelle
Avellaneda, Mario J.
Brouilly, Nicolas
Cezanne, Alice
Morales-Navarrete, Hernán
Perini, Enrico D.
Ferguson, Charles
Lupas, Andrei N.
Kalaidzidis, Yannis
Parton, Robert G.
Grill, Stephan W.
Zerial, Marino
author_sort Murray, David H.
collection PubMed
description An early step in intracellular transport is the selective recognition of a vesicle by its appropriate target membrane, a process regulated by Rab GTPases via the recruitment of tethering effectors1–4. Membrane tethering confers higher selectivity and efficiency to membrane fusion than the pairing of SNAREs alone5,6,7. Here, we addressed the mechanism whereby a tethered vesicle comes closer towards its target membrane for fusion by reconstituting an endosomal asymmetric tethering machinery consisting of the dimeric coiled-coil protein EEA16,7 recruited to phosphatidylinositol 3-phosphate membranes and binding vesicles harboring Rab5. Surprisingly, structural analysis revealed that Rab5:GTP induces an allosteric conformational change in EEA1, from extended to flexible and collapsed. Through dynamic analysis by optical tweezers we confirmed that EEA1 captures a vesicle at a distance corresponding to its extended conformation, and directly measured its flexibility and the forces induced during the tethering reaction. Expression of engineered EEA1 variants defective in the conformational change induced prominent clusters of tethered vesicles in vivo. Our results suggest a new mechanism in which Rab5 induces a change in flexibility of EEA1, generating an entropic collapse force that pulls the captured vesicle toward the target membrane to initiate docking and fusion.
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spelling pubmed-51426062017-02-24 An endosomal tether undergoes an entropic collapse to bring vesicles together Murray, David H. Jahnel, Marcus Lauer, Janelle Avellaneda, Mario J. Brouilly, Nicolas Cezanne, Alice Morales-Navarrete, Hernán Perini, Enrico D. Ferguson, Charles Lupas, Andrei N. Kalaidzidis, Yannis Parton, Robert G. Grill, Stephan W. Zerial, Marino Nature Article An early step in intracellular transport is the selective recognition of a vesicle by its appropriate target membrane, a process regulated by Rab GTPases via the recruitment of tethering effectors1–4. Membrane tethering confers higher selectivity and efficiency to membrane fusion than the pairing of SNAREs alone5,6,7. Here, we addressed the mechanism whereby a tethered vesicle comes closer towards its target membrane for fusion by reconstituting an endosomal asymmetric tethering machinery consisting of the dimeric coiled-coil protein EEA16,7 recruited to phosphatidylinositol 3-phosphate membranes and binding vesicles harboring Rab5. Surprisingly, structural analysis revealed that Rab5:GTP induces an allosteric conformational change in EEA1, from extended to flexible and collapsed. Through dynamic analysis by optical tweezers we confirmed that EEA1 captures a vesicle at a distance corresponding to its extended conformation, and directly measured its flexibility and the forces induced during the tethering reaction. Expression of engineered EEA1 variants defective in the conformational change induced prominent clusters of tethered vesicles in vivo. Our results suggest a new mechanism in which Rab5 induces a change in flexibility of EEA1, generating an entropic collapse force that pulls the captured vesicle toward the target membrane to initiate docking and fusion. 2016-08-24 2016-09-01 /pmc/articles/PMC5142606/ /pubmed/27556945 http://dx.doi.org/10.1038/nature19326 Text en Users may view, print, copy, and download text and data-mine the content in such documents, for the purposes of academic research, subject always to the full Conditions of use:http://www.nature.com/authors/editorial_policies/license.html#terms
spellingShingle Article
Murray, David H.
Jahnel, Marcus
Lauer, Janelle
Avellaneda, Mario J.
Brouilly, Nicolas
Cezanne, Alice
Morales-Navarrete, Hernán
Perini, Enrico D.
Ferguson, Charles
Lupas, Andrei N.
Kalaidzidis, Yannis
Parton, Robert G.
Grill, Stephan W.
Zerial, Marino
An endosomal tether undergoes an entropic collapse to bring vesicles together
title An endosomal tether undergoes an entropic collapse to bring vesicles together
title_full An endosomal tether undergoes an entropic collapse to bring vesicles together
title_fullStr An endosomal tether undergoes an entropic collapse to bring vesicles together
title_full_unstemmed An endosomal tether undergoes an entropic collapse to bring vesicles together
title_short An endosomal tether undergoes an entropic collapse to bring vesicles together
title_sort endosomal tether undergoes an entropic collapse to bring vesicles together
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5142606/
https://www.ncbi.nlm.nih.gov/pubmed/27556945
http://dx.doi.org/10.1038/nature19326
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