Calaxin stabilizes the docking of outer arm dyneins onto ciliary doublet microtubule in vertebrates
Abstract
Outer arm dynein (OAD) is the main force generator of ciliary beating. Although OAD loss is the most frequent cause of human primary ciliary dyskinesia, the docking mechanism of OAD onto the ciliary doublet microtubule (DMT) remains elusive in vertebrates. Here, we analyzed the functions of Calaxin/Efcab1 and Armc4, the two of five components of vertebrate OAD-DC (docking complex), using zebrafish spermatozoa and cryo-electron tomography. Mutation of armc4 caused complete loss of OAD, whereas mutation of calaxin caused only partial loss of OAD. Detailed structural analysis revealed that calaxin-/- OADs are tethered to DMT through DC components other than Calaxin, and that recombinant Calaxin can autonomously rescue the deficient DC structure and the OAD instability. Our data demonstrate the discrete roles of Calaxin and Armc4 in the OAD-DMT interaction, suggesting the stabilizing process of OAD docking onto DMT in vertebrates.
Data availability
The Source Data files contain the numerical data and raw gel images used to generate the figures.The maps generated in this study have been deposited in EMDB under the following accession numbers: EMD-34791, EMD-34792, EMD-34793, EMD-34794, EMD-34795, EMD-34796, EMD-34797, EMD-34798, EMD-34799, EMD-34800, EMD-34801, and EMD-34802.
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Doublet microtubule of zebrafish sperm axoneme, WTElectron Microscopy Data Bank, EMD-34791.
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Radial spoke 1 of zebrafish sperm axoneme, WTElectron Microscopy Data Bank, EMD-34792.
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Radial spoke 2 of zebrafish sperm axoneme, WTElectron Microscopy Data Bank, EMD-34793.
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Radial spoke 3 of zebrafish sperm axoneme, WTElectron Microscopy Data Bank, EMD-34794.
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Doublet microtubule of zebrafish sperm axoneme, calaxin-/-, OAD+ classElectron Microscopy Data Bank, EMD-34795.
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Doublet microtubule of zebrafish sperm axoneme, calaxin-/-, OAD- classElectron Microscopy Data Bank, EMD-34796.
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Doublet microtubule of zebrafish sperm axoneme, armc4-/-Electron Microscopy Data Bank, EMD-34797.
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Outer arm dynein of zebrafish sperm axoneme, WTElectron Microscopy Data Bank, EMD-34798.
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Outer arm dynein of zebrafish sperm axoneme, WT, refined focusing on the docking complexElectron Microscopy Data Bank, EMD-34799.
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Outer arm dynein of zebrafish sperm axoneme, WT, 1 mM calcium condition, refined focusing on the docking complexElectron Microscopy Data Bank, EMD-34800.
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Outer arm dynein of zebrafish sperm axoneme, calaxin-/-, refined focusing on the docking complexElectron Microscopy Data Bank, EMD-34801.
Article and author information
Author details
Funding
Japan Society for the Promotion of Science (KAKENHI Grant Number 16H02502)
- Masahide Kikkawa
Japan Society for the Promotion of Science (KAKENHI Grant Number 21H04762)
- Masahide Kikkawa
Japan Agency for Medical Research and Development (Grant Number JP22ama121002j001)
- Masahide Kikkawa
The funders had no role in study design, data collection and interpretation, or the decision to submit the work for publication.
Copyright
© 2023, Yamaguchi et al.
This article is distributed under the terms of the Creative Commons Attribution License permitting unrestricted use and redistribution provided that the original author and source are credited.
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