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Conformational diversity of dynactin sidearm and domain organization of its subunit p150

Dynactin is a principal regulator of the minus-end directed microtubule motor dynein. The sidearm of dynactin is essential for binding to microtubules and regulation of dynein activity. Although our understanding of the structure of the dynactin backbone (Arp1 rod) has greatly improved recently, str...

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Autores principales: Saito, Kei, Murayama, Takashi, Hata, Tomone, Kobayashi, Takuya, Shibata, Keitaro, Kazuno, Saiko, Fujimura, Tsutomu, Sakurai, Takashi, Toyoshima, Yoko Y.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: The American Society for Cell Biology 2020
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7353146/
https://www.ncbi.nlm.nih.gov/pubmed/32238103
http://dx.doi.org/10.1091/mbc.E20-01-0031
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author Saito, Kei
Murayama, Takashi
Hata, Tomone
Kobayashi, Takuya
Shibata, Keitaro
Kazuno, Saiko
Fujimura, Tsutomu
Sakurai, Takashi
Toyoshima, Yoko Y.
author_facet Saito, Kei
Murayama, Takashi
Hata, Tomone
Kobayashi, Takuya
Shibata, Keitaro
Kazuno, Saiko
Fujimura, Tsutomu
Sakurai, Takashi
Toyoshima, Yoko Y.
author_sort Saito, Kei
collection PubMed
description Dynactin is a principal regulator of the minus-end directed microtubule motor dynein. The sidearm of dynactin is essential for binding to microtubules and regulation of dynein activity. Although our understanding of the structure of the dynactin backbone (Arp1 rod) has greatly improved recently, structural details of the sidearm subcomplex remain elusive. Here, we report the flexible nature and diverse conformations of dynactin sidearm observed by electron microscopy. Using nanogold labeling and deletion mutant analysis, we determined the domain organization of the largest subunit p150 and discovered that its coiled-coil (CC1), dynein-binding domain, adopted either a folded or an extended form. Furthermore, the entire sidearm exhibited several characteristic forms, and the equilibrium among them depended on salt concentrations. These conformational diversities of the dynactin complex provide clues to understanding how it binds to microtubules and regulates dynein.
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spelling pubmed-73531462020-08-17 Conformational diversity of dynactin sidearm and domain organization of its subunit p150 Saito, Kei Murayama, Takashi Hata, Tomone Kobayashi, Takuya Shibata, Keitaro Kazuno, Saiko Fujimura, Tsutomu Sakurai, Takashi Toyoshima, Yoko Y. Mol Biol Cell Articles Dynactin is a principal regulator of the minus-end directed microtubule motor dynein. The sidearm of dynactin is essential for binding to microtubules and regulation of dynein activity. Although our understanding of the structure of the dynactin backbone (Arp1 rod) has greatly improved recently, structural details of the sidearm subcomplex remain elusive. Here, we report the flexible nature and diverse conformations of dynactin sidearm observed by electron microscopy. Using nanogold labeling and deletion mutant analysis, we determined the domain organization of the largest subunit p150 and discovered that its coiled-coil (CC1), dynein-binding domain, adopted either a folded or an extended form. Furthermore, the entire sidearm exhibited several characteristic forms, and the equilibrium among them depended on salt concentrations. These conformational diversities of the dynactin complex provide clues to understanding how it binds to microtubules and regulates dynein. The American Society for Cell Biology 2020-06-01 /pmc/articles/PMC7353146/ /pubmed/32238103 http://dx.doi.org/10.1091/mbc.E20-01-0031 Text en © 2020 Saito et al. “ASCB®,” “The American Society for Cell Biology®,” and “Molecular Biology of the Cell®” are registered trademarks of The American Society for Cell Biology. http://creativecommons.org/licenses/by-nc-sa/3.0 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.
spellingShingle Articles
Saito, Kei
Murayama, Takashi
Hata, Tomone
Kobayashi, Takuya
Shibata, Keitaro
Kazuno, Saiko
Fujimura, Tsutomu
Sakurai, Takashi
Toyoshima, Yoko Y.
Conformational diversity of dynactin sidearm and domain organization of its subunit p150
title Conformational diversity of dynactin sidearm and domain organization of its subunit p150
title_full Conformational diversity of dynactin sidearm and domain organization of its subunit p150
title_fullStr Conformational diversity of dynactin sidearm and domain organization of its subunit p150
title_full_unstemmed Conformational diversity of dynactin sidearm and domain organization of its subunit p150
title_short Conformational diversity of dynactin sidearm and domain organization of its subunit p150
title_sort conformational diversity of dynactin sidearm and domain organization of its subunit p150
topic Articles
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7353146/
https://www.ncbi.nlm.nih.gov/pubmed/32238103
http://dx.doi.org/10.1091/mbc.E20-01-0031
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