Far upstream element binding protein 2 interacts with enterovirus 71 internal ribosomal entry site and negatively regulates viral translation.
Far upstream element binding protein 2 interacts with enterovirus 71 internal ribosomal entry site and negatively regulates viral translation.
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DOI：
10.1093/nar/gkn901
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发表时间：
2009-01
影响因子：
14.9
通讯作者：
Shih SR
中科院分区：
文献类型：
--
作者：
Lin JY;Li ML;Shih SR
指导病毒蛋白翻译起始的内部核糖体进入位点（IRES）是肠病毒71型（EV71）的潜在药物靶点。内部起始的调控需要IRES反式作用因子（ITAFs）与内部核糖体进入位点的相互作用。采用生物素化rna亲和层析和蛋白质组学方法鉴定远上游元件（FUSE）结合蛋白2 （FBP2）为EV71的ITAF。FBP2与EV71 IRES的相互作用通过竞争分析和绘制病毒IRES和FBP2蛋白的关联位点得到证实。在EV71感染期间，FBP2富集于病毒复制发生的细胞质中，而在模拟感染细胞中，FBP2定位于细胞核中。在被EV71感染的fbp2敲低细胞中，病毒蛋白的合成增加。fbp2敲低细胞中的IRES活性高于阴性对照（NC） sirna处理细胞。另一方面，当FBP2在细胞中过表达时，IRES活性降低。本研究结果表明，FBP2是一种与EV71 IRES相互作用并负调控病毒翻译的新型ITAF。
An internal ribosomal entry site (IRES) that directs the initiation of viral protein translation is a potential drug target for enterovirus 71 (EV71). Regulation of internal initiation requires the interaction of IRES trans-acting factors (ITAFs) with the internal ribosomal entry site. Biotinylated RNA-affinity chromatography and proteomic approaches were employed to identify far upstream element (FUSE) binding protein 2 (FBP2) as an ITAF for EV71. The interactions of FBP2 with EV71 IRES were confirmed by competition assay and by mapping the association sites in both viral IRES and FBP2 protein. During EV71 infection, FBP2 was enriched in cytoplasm where viral replication occurs, whereas FBP2 was localized in the nucleus in mock-infected cells. The synthesis of viral proteins increased in FBP2-knockdown cells that were infected by EV71. IRES activity in FBP2-knockdown cells exceeded that in the negative control (NC) siRNA-treated cells. On the other hand, IRES activity decreased when FBP2 was over-expressed in the cells. Results of this study suggest that FBP2 is a novel ITAF that interacts with EV71 IRES and negatively regulates viral translation.