E3连接酶HUWE1与真核翻译起始因子eIF4E的相互作用

Interaction of E3 ligase HUWE1 and eukaryotic translation initiation factor eIF4E

为研究真核翻译起始因子eIF4E的调控,利用酵母双杂交系统从人骨髓cDNA文库筛选eIF4E相互作用蛋白质,核苷酸序列分析及同源性检索表明,其中一个约为0.4 kb的克隆与HUWE1(HECT,UBA和WWE domain containing 1,又称ARF-BP1,HECTH9,HUWE1)高度同源。细胞免疫共沉淀实验结果显示,在哺乳动物细胞水平能够检测到eIF4E蛋白与全长HUWE1的特异相互作用。利用构建好的含有HECT结构域的突变体进行相互作用位点研究,结果表明HUWE1的HECT结构域可以与eIF4E蛋白相互作用。

To explore the regulation of eIF4 E,we screened the protein interacting with eIF4 E from human cDNA library by using yeast two-hybrid system. Several clones interacting with eIF4 E were identified. One of them was homologous with HUWE1（HECT,UBA and WWE domain containing 1,also named as ARF-BP1,HECTH9 or HUWE1）. Cell co-immunoprecipitation showed that eIF4 E could bind to HUWE1 in mammalian cells. We also found that HUWE1 bearing the HECT domain is necessary for its association with eIF4 E.