1. Conformational diversity of dynactin sidearm and domain organization of its subunit p150
- Author
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Takashi Sakurai, Tomone Hata, Tsutomu Fujimura, Takuya Kobayashi, Saiko Kazuno, Yoko Y. Toyoshima, Keitaro Shibata, Takashi Murayama, and Kei Saito
- Subjects
Deletion mutant ,Protein subunit ,Dynein ,Molecular Conformation ,Dynein activity ,Regulator ,macromolecular substances ,Biology ,Dynactin Complex ,Microtubules ,Protein Domains ,Microtubule ,Amino Acid Sequence ,Molecular Biology ,urogenital system ,fungi ,Dyneins ,Articles ,Cell Biology ,Microscopy, Electron ,Cell Motility ,Dynactin ,Biophysics ,Microtubule-Associated Proteins ,Protein Binding - Abstract
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.
- Published
- 2020
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