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Mycalolide B dissociates dynactin and abolishes retrograde axonal transport of dense-core vesicles

Axonal transport is critical for maintaining synaptic transmission. Of interest, anterograde and retrograde axonal transport appear to be interdependent, as perturbing one directional motor often impairs movement in the opposite direction. Here live imaging of Drosophila and hippocampal neuron dense...

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Autores principales: Cavolo, Samantha L., Zhou, Chaoming, Ketcham, Stephanie A., Suzuki, Matthew M., Ukalovic, Kresimir, Silverman, Michael A., Schroer, Trina A., Levitan, Edwin S.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: The American Society for Cell Biology 2015
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4501363/
https://www.ncbi.nlm.nih.gov/pubmed/26023088
http://dx.doi.org/10.1091/mbc.E14-11-1564
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author Cavolo, Samantha L.
Zhou, Chaoming
Ketcham, Stephanie A.
Suzuki, Matthew M.
Ukalovic, Kresimir
Silverman, Michael A.
Schroer, Trina A.
Levitan, Edwin S.
author_facet Cavolo, Samantha L.
Zhou, Chaoming
Ketcham, Stephanie A.
Suzuki, Matthew M.
Ukalovic, Kresimir
Silverman, Michael A.
Schroer, Trina A.
Levitan, Edwin S.
author_sort Cavolo, Samantha L.
collection PubMed
description Axonal transport is critical for maintaining synaptic transmission. Of interest, anterograde and retrograde axonal transport appear to be interdependent, as perturbing one directional motor often impairs movement in the opposite direction. Here live imaging of Drosophila and hippocampal neuron dense-core vesicles (DCVs) containing a neuropeptide or brain-derived neurotrophic factor shows that the F-actin depolymerizing macrolide toxin mycalolide B (MB) rapidly and selectively abolishes retrograde, but not anterograde, transport in the axon and the nerve terminal. Latrunculin A does not mimic MB, demonstrating that F-actin depolymerization is not responsible for unidirectional transport inhibition. Given that dynactin initiates retrograde transport and that amino acid sequences implicated in macrolide toxin binding are found in the dynactin component actin-related protein 1, we examined dynactin integrity. Remarkably, cell extract and purified protein experiments show that MB induces disassembly of the dynactin complex. Thus imaging selective retrograde transport inhibition led to the discovery of a small-molecule dynactin disruptor. The rapid unidirectional inhibition by MB suggests that dynactin is absolutely required for retrograde DCV transport but does not directly facilitate ongoing anterograde DCV transport in the axon or nerve terminal. More generally, MB's effects bolster the conclusion that anterograde and retrograde axonal transport are not necessarily interdependent.
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spelling pubmed-45013632015-09-30 Mycalolide B dissociates dynactin and abolishes retrograde axonal transport of dense-core vesicles Cavolo, Samantha L. Zhou, Chaoming Ketcham, Stephanie A. Suzuki, Matthew M. Ukalovic, Kresimir Silverman, Michael A. Schroer, Trina A. Levitan, Edwin S. Mol Biol Cell Articles Axonal transport is critical for maintaining synaptic transmission. Of interest, anterograde and retrograde axonal transport appear to be interdependent, as perturbing one directional motor often impairs movement in the opposite direction. Here live imaging of Drosophila and hippocampal neuron dense-core vesicles (DCVs) containing a neuropeptide or brain-derived neurotrophic factor shows that the F-actin depolymerizing macrolide toxin mycalolide B (MB) rapidly and selectively abolishes retrograde, but not anterograde, transport in the axon and the nerve terminal. Latrunculin A does not mimic MB, demonstrating that F-actin depolymerization is not responsible for unidirectional transport inhibition. Given that dynactin initiates retrograde transport and that amino acid sequences implicated in macrolide toxin binding are found in the dynactin component actin-related protein 1, we examined dynactin integrity. Remarkably, cell extract and purified protein experiments show that MB induces disassembly of the dynactin complex. Thus imaging selective retrograde transport inhibition led to the discovery of a small-molecule dynactin disruptor. The rapid unidirectional inhibition by MB suggests that dynactin is absolutely required for retrograde DCV transport but does not directly facilitate ongoing anterograde DCV transport in the axon or nerve terminal. More generally, MB's effects bolster the conclusion that anterograde and retrograde axonal transport are not necessarily interdependent. The American Society for Cell Biology 2015-07-15 /pmc/articles/PMC4501363/ /pubmed/26023088 http://dx.doi.org/10.1091/mbc.E14-11-1564 Text en © 2015 Cavolo et al. This article is distributed by The American Society for Cell Biology under license from the author(s). Two months after publication it is available to the public under an Attribution–Noncommercial–Share Alike 3.0 Unported Creative Commons License (http://creativecommons.org/licenses/by-nc-sa/3.0). “ASCB®,” “The American Society for Cell Biology®,” and “Molecular Biology of the Cell®” are registered trademarks of The American Society for Cell Biology.
spellingShingle Articles
Cavolo, Samantha L.
Zhou, Chaoming
Ketcham, Stephanie A.
Suzuki, Matthew M.
Ukalovic, Kresimir
Silverman, Michael A.
Schroer, Trina A.
Levitan, Edwin S.
Mycalolide B dissociates dynactin and abolishes retrograde axonal transport of dense-core vesicles
title Mycalolide B dissociates dynactin and abolishes retrograde axonal transport of dense-core vesicles
title_full Mycalolide B dissociates dynactin and abolishes retrograde axonal transport of dense-core vesicles
title_fullStr Mycalolide B dissociates dynactin and abolishes retrograde axonal transport of dense-core vesicles
title_full_unstemmed Mycalolide B dissociates dynactin and abolishes retrograde axonal transport of dense-core vesicles
title_short Mycalolide B dissociates dynactin and abolishes retrograde axonal transport of dense-core vesicles
title_sort mycalolide b dissociates dynactin and abolishes retrograde axonal transport of dense-core vesicles
topic Articles
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4501363/
https://www.ncbi.nlm.nih.gov/pubmed/26023088
http://dx.doi.org/10.1091/mbc.E14-11-1564
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