CFAP70 Is a Novel Axoneme-Binding Protein That Localizes at the Base of the Outer Dynein Arm and Regulates Ciliary Motility.
CFAP70 Is a Novel Axoneme-Binding Protein That Localizes at the Base of the Outer Dynein Arm and Regulates Ciliary Motility.
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CFAP70是一种新型的轴突结合蛋白，该蛋白定位在外部动力蛋白臂的底部并调节纤毛运动。
DOI：
10.3390/cells7090124
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发表时间：
2018-08-29
期刊：
影响因子：
6
通讯作者：
Takeda S
中科院分区：
文献类型：
--
作者：
Shamoto N;Narita K;Kubo T;Oda T;Takeda S
在本研究中，我们对小鼠纤毛相关蛋白候选CFAP70进行了表征。由于该蛋白具有一簇四肽重复（TPR）结构域，就像许多纤束内运输（IFT）复合物的成分一样，我们研究了纤毛靶向和/或定位中特别感兴趣的结构域功能。RT-PCR和免疫组化检测显示，CFAP70与纤毛和鞭毛的运动有关。从猪气管纤毛中逐步提取的蛋白质表明，CFAP70与纤毛轴突紧密结合。对表达CFAP70片段的室管膜进行荧光显微镜观察发现，在纤毛定位中，n端比含有TPR结构域的c端更重要。当CFAP70在培养的小鼠室管膜中被敲除时，观察到纤毛跳动频率的减少。与这些观察结果一致的是，缺乏CFAP70同源物的衣藻突变体FAP70表现出外动力蛋白臂（ODA）活性缺陷和鞭毛运动性降低。冷冻电子断层扫描显示，FAP70的n端稳定地位于ODA的底部。这些结果表明，CFAP70是运动纤毛和鞭毛中ODA的一个新的调控成分，并且n端对其纤毛定位很重要。
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.
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影响因子：
3.3
作者：
Kubo T;Hou Y;Cochran DA;Witman GB;Oda T
通讯作者：
Oda T
影响因子：
3.3
作者：
Kubo T;Yanagisawa HA;Liu Z;Shibuya R;Hirono M;Kamiya R
通讯作者：
Kamiya R
影响因子：
3
作者：
Metlagel, Zoltan;Kikkawa, Yayoi S.;Kikkawa, Masahide
通讯作者：
Kikkawa, Masahide
影响因子：
3.3
作者：
Kubo T;Oda T
通讯作者：
Oda T
影响因子：
56.9
作者：
Jinek, Martin;Chylinski, Krzysztof;Charpentier, Emmanuelle
通讯作者：
Charpentier, Emmanuelle