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The carboxyl-terminal region of SDCCAG8 comprises a functional module essential for cilia formation as well as organ development and homeostasis

堤, 崚太郞 大阪大学 DOI:10.18910/87841

2022.03.24

概要

繊毛は細胞の表面に突出した微小管の軸を持つ細胞小器官であり、生物の発生や恒常性維持に重要な役割を担っている。ヒトにおける繊毛の機能不全は、水頭症、網膜色素変性症、嚢胞腎、肥満、糖尿病、不妊、内臓逆位、難聴、多指症、短肢症などの症状が見られる「繊毛病」と呼ばれる一群の疾患を引き起こす。しかし、繊毛病の根本的な治療法はまだ確立されておらず、繊毛病の発症・病態メカニズムについてもまだ不明な点が多く残っている。

当研究室の先行研究により、互いに相同性の高いセリンスレオニンキナーゼであるIntestinal cell kinase(Ick)とMale germ cell-associated kinase(Mak)が、繊毛内タンパク質輸送と繊毛の長さを制御することが明らかとなった。ヒトICK遺伝子のミスセンス変異は、致死となる繊毛病の原因であることが報告されている。また、ヒトMAK遺伝子の変異は網膜色素変性症の原因となることが知られている。我々は、繊毛病の分子病態メカニズムを明らかにするため、Ickと相互作用する分子をスクリーニングにより探索し、繊毛病原因遺伝子として知られるSerologically defined colon cancer antigen8(Sdccag8)を候補因子として同定した。免疫沈降法によって、IckとMakはSdccag8のC末端領域(Sdccag8-C)と相互作用することが明らかとなった。ヒトSDCCAG8のC末端領域を欠損させると考えられる短縮型変異が、網膜や腎臓の変性などの様々な器官の異常と関連すると報告されている。私は、この知見に基づいてSdccag8のC末端領域に着目し、培養細胞を用いた実験により、Sdccag8-CがSdccag8の基底小体への局在と繊毛形成に必要であることを明らかにした。さらに、in vivoにおけるSdccag8-Cの役割を明らかにするために、ゲノム編集を用いてSdccag8-Cが欠失するように終始コドンをノックインしたSdccag8部分欠失(Sdccag8ΔC/ΔC)マウスを作製した。Sdccag8ΔC/ΔCマウスを解析したところ、Sdccag8ΔC/ΔCマウスは、口蓋裂、多指、網膜と腎臓の変性、精子形成異常といった繊毛病様の表現型を示した。また、そのSdccag8ΔC/ΔCマウスの表現型は、Ick欠損マウスやMak欠損マウスの表現型と一部類似していた。以上の結果から、Sdccag8-CはSdccag8の基底小体への局在と繊毛形成に必要であること、Sdccag8はIck、Makと機能的に相互作用していることが示唆された。本研究により、繊毛病原因タンパク質の相互作用ネットワークと繊毛病の病態メカニズムの一端が解明されたと考えられる。

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