Ota Yu, Chinen Takumi, Yoshida Keisuke, Kudo Shun, Nagumo Yoko, Shiwa Yuh, Yamada Ryosuke, Umihara Hirotatsu, Iwasaki Kotaro, Masumoto Hiroshi, Yokoshima Satoshi, Yoshikawa Hirofumi, Fukuyama Tohru, Kobayashi Junichi, Usui Takeo
Faculty of Life and Environmental Sciences, University of Tsukuba, 1-1-1 Tennodai, Tsukuba, Ibaraki, 305-8572, Japan.
NODAI Genome Research Center, Tokyo University of Agriculture, 1-1-1 Sakuragaoka, Setagaya-ku, Tokyo, 156-8502, Japan.
Chembiochem. 2016 Sep 2;17(17):1616-20. doi: 10.1002/cbic.201600075. Epub 2016 Jul 22.
Eudistomin C (EudiC), a natural product, shows potent antitumor and antiviral activities, but the target molecule and the mechanism of action remain to be revealed. Here, we show that the 40S ribosome is the target in EudiC cytotoxicity. We isolated EudiC-resistant mutants from a multidrug-sensitive yeast strain, and a genetic analysis classified these YER (yeast EudiC resistance) mutants into three complementation groups. A genome-wide study revealed that the YER1-6 mutation is in the uS11 gene (RPS14A). Biotinylated EudiC pulled down Rps14p-containing complexes from 40S and 80S ribosomes, but not from the 60S ribosome. EudiC strongly inhibited translation of the wild-type strain but not of YER1-6 in cells and in vitro. These results indicate that EudiC is a protein synthesis inhibitor targeting the uS11-containing ribosomal subunit, and shows cytotoxicity by inhibiting protein translation.
优地斯明 C（EudiC）是一种天然产物，具有强大的抗肿瘤和抗病毒活性，但其靶分子和作用机制仍有待揭示。在此，我们表明40S核糖体是EudiC细胞毒性作用的靶点。我们从一株多药敏感酵母菌株中分离出EudiC抗性突变体，基因分析将这些YER（酵母EudiC抗性）突变体分为三个互补组。全基因组研究表明，YER1 - 6突变位于uS11基因（RPS14A）中。生物素化的EudiC从40S和80S核糖体中拉下了含Rps14p的复合物，但未从60S核糖体中拉下。EudiC在细胞内和体外均强烈抑制野生型菌株的翻译，但不抑制YER1 - 6的翻译。这些结果表明，EudiC是一种靶向含uS11核糖体亚基的蛋白质合成抑制剂，通过抑制蛋白质翻译表现出细胞毒性。
