Tomita Takeshi, Ieguchi Katsuaki, Coin Frédéric, Kato Yasuhiro, Kikuchi Haruhisa, Oshima Yoshiteru, Kurata Shoichiro, Maru Yoshiro
Department of Pharmacology, Tokyo Women's Medical University, 8-1 Kawada-cho, Shinjuku-ku, Tokyo, Japan.
Graduate School of Pharmaceutical Sciences, Tohoku University, Sendai, Japan.
PLoS One. 2014 Sep 30;9(9):e108957. doi: 10.1371/journal.pone.0108957. eCollection 2014.
Celastramycin A, a small molecule that inhibits the production of antibacterial peptides in an ex vivo culture system of Drosophila, suppresses the TNFα-mediated induction of IL-8 in mammalian cells. To understand its molecular mechanism, we examined Celastramycin A binding proteins and investigated their biological functions. Our screening and subsequent pull-down assay revealed ZFC3H1 (also known as CCDC131 or CSRC2), an uncharacterized zinc finger protein, as a Celastramycin A binding protein. The knockdown of ZFC3H1 reduced IL-8 expression levels in the TNFα-stimulated lung carcinoma cell line, LU99, and UV-irradiated HeLa cells. Based on reporter assay results, we concluded that ZFC3H1 participates in the transcriptional activation of IL-8. The findings of our UV-irradiation experiments implied that ZFC3H1 may indirectly interact with ERCC1 in an activated DNA repair complex. Thus, we designated ZFC3H1 as a mammalian target of Celastramycin A (mTOC).
南蛇藤霉素A是一种小分子化合物，它在果蝇的体外培养系统中抑制抗菌肽的产生，在哺乳动物细胞中可抑制TNFα介导的IL-8诱导。为了解其分子机制，我们检测了南蛇藤霉素A结合蛋白并研究了它们的生物学功能。我们的筛选及随后的下拉分析显示，一种未被鉴定的锌指蛋白ZFC3H1（也称为CCDC131或CSRC2）是南蛇藤霉素A结合蛋白。敲低ZFC3H1可降低TNFα刺激的肺癌细胞系LU99和紫外线照射的HeLa细胞中IL-8的表达水平。基于报告基因检测结果，我们得出结论，ZFC3H1参与IL-8的转录激活。我们紫外线照射实验的结果表明，ZFC3H1可能在活化的DNA修复复合物中与ERCC1间接相互作用。因此，我们将ZFC3H1指定为南蛇藤霉素A的哺乳动物靶点（mTOC）。
