ケジカルゲノミクスの手法でオートファジー制御活性を定量的に測定した。その結果を階層的クラスタリングによって解析することで, がんと神経細胞におけるオートファジーの制御機構の違いを明らかにした。オートファジーを促進することで, たんぱく質凝集を阻害するSO286はSOBP1と結合し, SOBPをノックダウンすることでSO286の作用が阻害された。このことから, SOBP1がSO286の作用に関与していることが示唆された。オートファジー細胞死誘導物質の探索をおこなった結果, 放線菌2142A-36株を大量培養し, 単離精製, 構造解析した結果, Cholest-5-ene-3b,7a-diolを取得した。
We constructed GFP-LC3-RFP-expressing cells, and using the cells, we analyzed the diversity and consistency of regulatory signaling in autophagy. The effects of more than 200 small molecular compounds were assessed quantitatively by autophagy regulatory system in cancer cells and neuronal cells. Hierarchical clustering was performed on the subsequent autophagy inhibition profile of the compounds in each cell type. The result was that hierarchical clustering accurately classified the compounds according to their targets. And we distinguished between common and cell type-specific signals responsible for autophagy. SO286, which increased autophagy flux, inhibited the Aggresome formation. SO286 was found to bind SOBP, and by knockdown of SOBP, SO286 failed to inhibit Aggresome formation. This result indicated that SO286 functioned SOBP-dependent manner. We searched for the compounds that induce autophagic cell death, and we isolated Cholest-5-ene-3b,7a-diol from microbial origin.
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