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Cells 2018, 7(9), 124; https://doi.org/10.3390/cells7090124

CFAP70 Is a Novel Axoneme-Binding Protein That Localizes at the Base of the Outer Dynein Arm and Regulates Ciliary Motility

1
Department of Anatomy and Cell Biology, Interdisciplinary Graduate School, University of Yamanashi, Chuo, Yamanashi 409-3898, Japan
2
Department of Anatomy and Structural Biology, Interdisciplinary Graduate School, University of Yamanashi, Chuo, Yamanashi 409-3898, Japan
These authors contributed equally to this work.
*
Authors to whom correspondence should be addressed.
Received: 3 August 2018 / Revised: 24 August 2018 / Accepted: 28 August 2018 / Published: 29 August 2018
(This article belongs to the Special Issue Cilia and Flagella: Structure, Function and Beyond)
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Abstract

In the present study, we characterized CFAP70, a candidate of cilia-related protein in mice. As this protein has a cluster of tetratricopeptide repeat (TPR) domains like many components of the intraflagellar transport (IFT) complex, we investigated the domain functions of particular interest in ciliary targeting and/or localization. RT-PCR and immunohistochemistry of various mouse tissues demonstrated the association of CFAP70 with motile cilia and flagella. A stepwise extraction of proteins from swine tracheal cilia showed that CFAP70 bound tightly to the ciliary axoneme. Fluorescence microscopy of the cultured ependyma expressing fragments of CFAP70 demonstrated that the N-terminus rather than the C-terminus with the TPR domains was more important for the ciliary localization. When CFAP70 was knocked down in cultured mouse ependyma, reductions in cilia beating frequency were observed. Consistent with these observations, a Chlamydomonas mutant lacking the CFAP70 homolog, FAP70, showed defects in outer dynein arm (ODA) activity and a reduction in flagellar motility. Cryo-electron tomography revealed that the N-terminus of FAP70 resided stably at the base of the ODA. These results demonstrated that CFAP70 is a novel regulatory component of the ODA in motile cilia and flagella, and that the N-terminus is important for its ciliary localization. View Full-Text
Keywords: tetratricopeptide repeat containing protein; outer dynein arm; cryo-electron tomography tetratricopeptide repeat containing protein; outer dynein arm; cryo-electron tomography
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Shamoto, N.; Narita, K.; Kubo, T.; Oda, T.; Takeda, S. CFAP70 Is a Novel Axoneme-Binding Protein That Localizes at the Base of the Outer Dynein Arm and Regulates Ciliary Motility. Cells 2018, 7, 124.

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