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

RNA 结合和代谢：阐明糖酵解酶在转录后基因调控中的作用

• 批准号: BB/N008820/1
• 负责人: Andre Gerber
• 金额: $ 51.51万
• 依托单位: University of Surrey
• 依托单位国家: 英国
• 项目类别: Research Grant
• 财政年份: 2016
• 资助国家: 英国
• 起止时间: 2016 至 无数据
• 项目状态: 已结题
• 来源: https://gtr.ukri.org/projects?ref=BB%2FN008820%2F1
• 关键词: RNA; Binding; Metabolism; Elucidating; Role

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.

细胞是生物体的最小单位。每个生物体的所有细胞都包含相同遗传信息的副本，这些信息以DNA的形式组织成基因。在基因表达的过程中，DNA被复制到一种名为RNA的中间分子，然后它可以作为合成蛋白质的模板。蛋白质定义了有机体中每个细胞的形状和功能。从物理上讲，RNA被蛋白质覆盖，也就是所谓的RNA结合蛋白。这些蛋白质可以移除或重新排列部分RNA，将其储存或传递到细胞内的特定位置，并最终将其降解。它们还控制信使RNAs(mRNAs；指的是编码蛋白质的一类RNAs)何时和如何翻译成蛋白质。如果RNA结合蛋白不能正常工作，就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对细胞中与mRNAs相互作用的所有RNA结合蛋白进行分类。除了检测以前验证的RNA结合蛋白外，我们还发现许多具有其他成熟功能的蛋白质，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，它们提供和控制维持细胞存活所需的能量流动。虽然以前已经描述了酶与RNA结合的个别例子，但发现一种重要的能量产生途径，即糖酵解的大多数甚至所有酶都与酵母中的RNA相互作用，这一发现引起了我们的兴趣。在糖酵解过程中，葡萄糖通过一条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都高度保守，并受到严格控制。在酵母中，糖酵解是从葡萄糖中产生乙醇的关键，这是酵母的一个特征，几个世纪以来，酵母一直被用于生产葡萄酒、啤酒和其他商品。在人类中，该途径在癌细胞中高度活跃，因此为开发癌症治疗的新策略提供了一个靶点。我们发现这一重要途径中的所有酶都与RNA结合，对此我们很感兴趣，我们希望了解这些相互作用的基础和功能。因此，我们建议全面鉴定这一途径的酶的RNA靶标。然后，我们将调查它们在哪里以及如何与RNA结合，特别是取消细胞中的mRNA结合部位，看看它是否影响特定mRNAs的命运。同样，我们将测量取消这种结合是否会对mRNAs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，我们希望发现以前未知的RNA调控和燃料电池化学反应之间的联系。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。

项目成果

Oxidative stress induces coordinated remodeling of RNA-enzyme interactions.

• DOI: 10.1016/j.isci.2021.102753
• 发表时间: 2021-07-23
• 期刊: iScience
• 影响因子: 5.8
• 作者: Matia-González AM;Jabre I;Laing EE;Gerber AP
• 通讯作者: Gerber AP

RNA-Centric Approaches to Profile the RNA-Protein Interaction Landscape on Selected RNAs.

• DOI: 10.3390/ncrna7010011
• 发表时间: 2021-02-15
• 期刊: Non-coding RNA
• 影响因子: 4.3
• 作者: Gerber AP

Mirroring the multifaceted role of RNA and its partners in gene expression.

反映 RNA 及其伙伴在基因表达中的多方面作用。

• DOI: 10.1002/1873-3468.13230
• 发表时间: 2018
• 期刊: FEBS letters
• 影响因子: 3.5
• 作者: Zavolan M

Biochemical approach for isolation of polyadenylated RNAs with bound proteins from yeast.

• DOI: 10.1016/j.xpro.2021.100929
• 发表时间: 2021-12-17
• 期刊: STAR protocols
• 作者: Matia-González AM;Jabre I;Gerber AP
• 通讯作者: Gerber AP