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

Cells
Noritoshi ShamotoSen Takeda

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 c...Continue Reading

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Citations

Jan 18, 2020·Human Genetics·Aminata TouréCharles Coutton
Jun 20, 2020·Journal of Human Genetics·Yanwei ShaShaobin Lin
Sep 12, 2020·Cellular and Molecular Life Sciences : CMLS·Lance Lee, Lawrence E Ostrowski
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Dec 12, 2020·Cancer Science·Sohei KobayashiKazuyuki Matsushita
Nov 17, 2020·Best Practice & Research. Clinical Endocrinology & Metabolism·Chaofeng TuYue-Qiu Tan
Apr 10, 2021·G3 : Genes - Genomes - Genetics·Irina SizovaPeter Hegemann

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Datasets Mentioned

BETA
EMD-9618

Methods Mentioned

BETA
affinity purification
PCR
electron microscopy
fluorescence microscopy

Software Mentioned

psipred
ImageJ
Fluoview
TI Workbench

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