ASCB/EMBO 2019 meeting 参加報告

社本 憲俊

医学部医学科 2014年入学

2019年12月7-11日 米国ワシントンD.C.

2019126日よりワシントンD.C.にて開催されたASCB (American Society for Cell Biology) に参加しました。この度は、このような素晴らしい機会をくださった竹田先生や成田先生を始めとする解剖学講座細胞生物学教室の先生方に、この場をお借りして感謝申し上げます。初めての海外における学会発表あり、とても貴重な経験を積むことができました。

発表はポスター発表を行い、1時間半の発表時間の中で、9人もの方が聞きに来てくださりました。多くの質問やアイデアをいただき、中には論文を読んでくださっている方もいました。また、ChlamydomonasC. elegans に詳しい方々とは、FAP70の局在について深い意見を交わすことができました。今回の経験を糧として、基礎・臨床の両方において、これからも努力していきたいと思います。以下に抄録を記します。

Cilia- and flagella-associated protein 70 (CFAP70) is a 150 kDa protein discovered by our transcriptome analysis in mice. Little is known about this protein, except for the tetratricopeptide repeat (TPR) domains concentrated on the C-terminus, which is known to mediate protein-protein interactions, and present in many components of the intraflagellar transport (IFT) complex. We characterized the mouse CFAP70 and Chlamydomonas homolog FAP70 with particular interests on the ciliary localization and function. RT-PCR of various mouse tissues demonstrated the association of CFAP70 with motile cilia and flagella, and the immunohistochemistry showed a staining pattern being different from the punctate signals of IFT subunits. A stepwise extraction of proteins from porcine 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 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. However, there are no obvious changes in the FAP70-null mutant axoneme. These results collectively demonstrate that CFAP70 is a novel regulatory component of the ODA in motile cilia and flagella. Furthermore, binding of CFAP70 to the axoneme takes places at the N-terminus being independent of TPR domain.



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