RNA Binding and Metabolism: Elucidating the Role of Glycolytic Enzymes in Posttranscriptional Gene Regulation

细胞是有机体的最小单位。每个生物体的所有细胞都包含相同遗传信息的副本，这些遗传信息以DNA的形式组织成基因。在基因表达过程中，DNA被复制成一种称为RNA的中间分子，然后RNA可以作为蛋白质合成的模板。蛋白质定义了生物体每个细胞的形状和功能。在物理上，RNA被蛋白质覆盖，即所谓的RNA结合蛋白。这些蛋白质可以移除或重新排列RNA的一部分，将其储存或运送到细胞内的特定位置，并最终降解它。它们还控制信使RNA（mRNA;指编码蛋白质的RNA类）何时以及如何翻译成蛋白质。如果RNA结合蛋白不能正常工作，它可能导致细胞功能障碍，最终导致疾病。由于它们的巨大重要性，我们和其他研究人员使用了一种新的实验方法来编目细胞中与mRNA相互作用的所有RNA结合蛋白。除了检测先前验证的RNA结合蛋白外，我们发现许多具有其他功能的蛋白质，如酶，也能够与RNA相互作用。酶是执行基本化学反应的蛋白质;例如，它们提供并控制保持细胞存活所需的能量流。虽然之前已经描述了与RNA结合的酶的个别例子，但发现大多数甚至所有重要的能量产生途径（称为糖酵解）的酶与酵母中的RNA相互作用引起了我们的兴趣。在糖酵解过程中，葡萄糖通过涉及几个连续步骤的途径转化为另一种化学物质。在这个过程中，产生了能量。这一重要途径在所有生物体中高度保守，并受到严格控制。在酵母中，糖酵解对于从葡萄糖生成乙醇是必不可少的，这是酵母的一个特征，几个世纪以来一直用于生产葡萄酒，啤酒和其他商品。在人类中，该途径在癌细胞中高度活跃，因此为癌症治疗新策略的开发提供了靶点。我们的研究结果是，在这一重要途径中的所有酶都与RNA结合，这引起了我们的兴趣，我们希望了解这些相互作用的基础和功能。因此，我们建议全面确定该途径酶的RNA靶点。然后，我们将研究它们在哪里以及如何与RNA结合，并特异性地消除细胞中的mRNA结合位点，以观察它是否影响特定mRNA的命运。同样，我们将测量取消这种结合是否会对mRNA编码的蛋白质的活性和相关途径产生影响。通过我们的研究，我们希望发现RNA调节和燃料电池化学反应之间以前未被认识的联系。这些知识可能会对我们社会的各个重要方面产生影响，从食品生产到新的癌症治疗方法的开发。

A cell is the smallest unit of an organism. All cells of every organism contain a copy of the same genetic information, which is organised into genes in the form of DNA. During the process of gene expression, DNA is copied to an intermediate molecule called RNA, which can then serve as a template for the synthesis of proteins. Proteins define the shape and function of each cell of the organism. Physically, RNAs are covered by proteins, so-called RNA-binding proteins. These proteins can remove or rearrange parts of the RNA, store, or deliver it to particular locations within the cell, and ultimately degrade it. They also control when and how messenger RNAs (mRNAs; refers to the class of RNAs that encode proteins) are translated into proteins. If an RNA-binding protein does not work properly, it can lead to malfunction of the cell and ultimately to disease. Due to their tremendous importance, we and other researchers have used a new experimental approach to catalogue all of the RNA-binding proteins that interact with mRNAs in cells. Besides detecting previously validated RNA-binding proteins, we found that many proteins with other well-established functions, such as enzymes, are also able to interact with RNA. Enzymes are proteins that perform essential chemical reactions; for instance, they provide and control the flow of energy required to keep cells alive. Whilst individual examples of enzymes binding to RNA have been characterised previously, the finding that most or even all enzymes of an important energy-generating pathway, termed glycolysis, interact with RNAs in yeast intrigued us. During glycolysis, glucose is transformed via a pathway involving several sequential steps, into another chemical. During this process, energy is generated. This important pathway is highly conserved in all organisms and is tightly controlled. In yeast, glycolysis is essential for the generation of ethanol from glucose, a feature of yeast that has been used for centuries for the production of wine, beer and other goods. In humans, the pathway is highly active in cancer cells and thus, provides a target for the development of new strategies for cancer treatment. Intrigued by our findings that all of the enzymes in this essential pathway bind to RNA, we wish to understand both the basis and the function of these interactions. Therefore, we propose to comprehensively identify the RNA targets for the enzymes of this pathway. We will then investigate where and how they bind to the RNA, and specifically abrogate mRNA-binding sites in cells, to see whether it affects the fate of particular mRNAs. Likewise, we will measure whether abolishing this binding has an effect on the activity of the protein encoded by the mRNAs and the associated pathway.With our research, we hope to discover previously unrecognised connections between RNA regulation and the chemical reactions that fuel cells. This knowledge is likely to have impact on diverse important aspects of our society, from food-production, to the development of new cancer treatments.