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首页> 外文期刊>Molecular and Cellular Biology >Novel Regulatory Factors Interacting with the Promoter of the Gene Encoding the mRNA Cap Binding Protein (eIF4E) and Their Function in Growth Regulation
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Novel Regulatory Factors Interacting with the Promoter of the Gene Encoding the mRNA Cap Binding Protein (eIF4E) and Their Function in Growth Regulation

机译：与编码mRNA帽结合蛋白（eIF4E）基因的启动子相互作用的新型调控因子及其在生长调节中的功能

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Regulation of the mRNA cap binding protein (eIF4E) is critical to the control of cellular proliferation since this protein is the rate-limiting factor in translation initiation and transforms fibroblasts and since eIF4E mutants arrest budding yeast in the G1 phase of the cell cycle (cdc33). We previously demonstrated regulation of eIF4E by altered transcription of its mRNA in serum-stimulated fibroblasts and in response to c-myc. To identify additional factors regulating eIF4E transcription, we used linker-scanning constructs to characterize sites in the promoter of the eIF4E gene required for its expression. Promoter activity was dependent on sites at ?5, ?25, ?45, and ?75; the site at ?75 included a previously described myc box. Electrophoretic mobility shift assays identified DNA-protein interactions at ?25 and revealed a binding site (TTACCCCCCCTT) that is unique to the eIF4E promoter. Proteins of 68 and 97 kDa bound this site in UV cross-linking and Southwestern experiments. Levels of 4E regulatory factor activities correlated with c-Myc levels, eIF4E expression levels, and protein synthesis in differentiating U937 and HL60 cells, suggesting that these activities may function to regulate protein synthesis rates during differentiation. Since the eIF4E promoter lacked typical TATA and initiator elements, further studies of this novel initiator-homologous element should provide insights into mechanisms of transcription initiation and growth regulation.
机译：mRNA帽结合蛋白（eIF4E）的调节对于控制细胞增殖至关重要，因为该蛋白是翻译起始和转化成纤维细胞的速率限制因子，并且因为eIF4E突变体将G 1 细胞周期的阶段（ cdc33 ）。我们先前证明了eIF4E的调节是通过改变其在血清刺激的成纤维细胞中的mRNA转录以及响应c- myc 来实现的。为了确定调节eIF4E转录的其他因素，我们使用了连接子扫描构建体来表征其表达所需的eIF4E基因启动子中的位点。启动子活性取决于在？5，？25，？45和？75的位点; 75欧元处的站点包括一个先前描述的 myc 框。电泳迁移率变动分析鉴定了在25处的DNA-蛋白质相互作用，并揭示了eIF4E启动子特有的结合位点（TTACCCCCCCTT）。在紫外线交联和西南实验中，68 kDa和97 kDa的蛋白质结合了该位点。在分化的U937和HL60细胞中，4E调节因子活性的水平与c-Myc水平，eIF4E表达水平和蛋白质合成相关，这表明这些活性可能在分化过程中起到调节蛋白质合成速率的作用。由于eIF4E启动子缺少典型的TATA和启动子元件，因此对该新型启动子-同源元件的进一步研究应提供有关转录启动和生长调控机制的见解。

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