Translational Control by Human Pumilio Proteins

项目摘要 人RNA结合蛋白，R11和R12，是哺乳动物发育所必需的， 功能障碍与多种人类疾病有关，包括发育缺陷，神经障碍， 不孕症癌症和线粒体功能障碍这些重要的职能迫使我们的总体目标是 发现BMP1和BMP2如何控制遗传信息从基因到mRNA再到蛋白质的流动， 它们所调控的全部基因在人类细胞中，BMP1和BMP2通过识别 Pumilio响应元件（PRE）。先前的研究表明， 通过募集RNA降解酶促进数百种含有PRE的mRNA的降解。现在 然而，很明显，这种机制只代表了一种类型的神经网络介导的调节结果。数千 的mRNA被β 1和β 2结合，但不被降解。因此，现在有必要确定如何 β 1和β 2控制所有靶mRNA的命运。由此产生的数据将提供一个全面的看法， 在生物学和发病机制中的调节作用。 我们认为人的BMP1和BMP2通过抑制翻译过程来抑制许多靶mRNA。 这一假设得到了多个基因在蛋白质水平上被BMP1和BMP2抑制的例子的支持 在没有mRNA降解的情况下的丰度。这种翻译抑制的机制和普遍性 不明此外，我们的数据表明，对于某些基因，β-内酰胺酶介导的翻译抑制可以 与RNA降解协同作用以比单独的任一过程更大程度地调节基因表达。 RNA分子形成影响其功能和命运的结构。虽然生化证据 表明RNA结构可以调节RNA-PRE相互作用，但其在体内的作用尚不清楚。事实上， 人类细胞中mRNA的RNA结构信息总体上缺乏，这限制了我们对 这种结构是如何通过RNA结合蛋白（如RNA结合蛋白1和RNA结合蛋白2）影响基因调控的。 这项研究旨在确定BMP1和BMP2如何抑制翻译，并确定BMP1和BMP2在翻译中的作用。 翻译调节因子，其是α 1和β 2活性所必需的。人类mRNA的结构将 并分析其对mRNA相互作用和调控网络的影响。通过整合 这些新的数据与现有的知识，其中mRNA的结合和降解的BMP1和2，我们将开发一个 全面了解这一关键的遗传调控网络。发现完整的监管网络， EST1&2将为它们如何控制基因表达以调节正常的生物过程提供新的见解。 此外，这些知识将有助于阐明他们的功能障碍如何导致神经变性等疾病 和癌症

Project Summary The human RNA-binding proteins, PUM1 and PUM2, are essential for mammalian development and their dysfunction is linked to multiple human diseases including developmental defects, neurological disorders, infertility, cancers, and mitochondrial dysfunction. These important functions compel our overall objective to discover how PUM1&2 control the flow of genetic information from gene to mRNA to protein and to identify the full repertoire of genes that they regulate. PUM1&2 bind to thousands of mRNAs in human cells by recognizing an RNA sequence called the Pumilio Response Element (PRE). Previous research showed that PUM1&2 promote degradation of hundreds of these PRE-containing mRNAs by recruiting RNA decay enzymes. It is now clear, however, that this mechanism represents only one type of PUM-mediated regulatory outcome. Thousands of mRNAs are bound by PUM1&2 but are not degraded. Therefore, it is now necessary to determine how PUM1&2 control the fate of all target mRNAs. The resulting data will provide a comprehensive view of their regulatory roles in biology and pathogenesis. We propose that human PUM1&2 repress many target mRNAs by inhibiting the process of translation. This hypothesis is supported by multiple examples of genes that are repressed by PUM1&2 at the level of protein abundance in the absence of mRNA degradation. The mechanism and prevalence of this translational inhibition is unknown. In addition, our data indicate that for some genes PUM-mediated translational inhibition can synergize with RNA degradation to regulate gene expression to a larger extent than either process alone. RNA molecules form structures that influence their function and fate. While biochemical evidence indicates that RNA structure can modulate PUM-PRE interactions, its effect in vivo remains unknown. In fact, there is an overall lack of RNA structural information of mRNAs in human cells that limits our understanding of how that structure influences gene regulation by RNA-binding proteins like PUM1&2. The proposed research seeks to determine how PUM1&2 inhibit translation and to identify the translational regulatory factors that are necessary for PUM1&2 activity. The structure of human mRNAs will be determined and its effect on PUM-mRNA interactions and regulatory network will be analyzed. By integrating this new data with existing knowledge of which mRNAs are bound and degraded by PUM1&2, we will develop a comprehensive understanding of this key genetic regulatory network. Discovery of the full regulatory network of PUM1&2 will provide new insights into how they control gene expression to regulate normal biological processes. Moreover, this knowledge will help elucidate how their dysfunction leads to diseases such as neurodegeneration and cancer.