Small Molecule Effectors of Eukaryotic Translation Initiation Site Selection

真核翻译起始位点选择的小分子效应器

• 批准号: 7467404
• 负责人: JON R. LORSCH
• 金额: $ 24.11万
• 依托单位: JOHNS HOPKINS UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2007
• 资助国家: 美国
• 起止时间: 2007-07-15 至 2010-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7467404
• 关键词: Address; Affect; Amino Acid Sequence; Amino Acyl Transfer RNA; Aminoglycosides; Anti-Bacterial Agents; Antifungal Agents; Area; Bacteria; Biological Assay; Cells; Chemicals; Class; Codon Nucleotides; Collection; Complex; Depth; Development; Diabetes Insipidus; Disease; Etiology; Eukaryota; Eukaryotic Cell; Event; Future; Gene Expression; Gene Expression Regulation; Genes; Heart; Hereditary Disease; Housing; In Vitro; Infection; Initiator Codon; Instruction; Lead; Luciferases; Malignant Neoplasms; Mammalian Cell; Mammals; Messenger RNA; Molecular; Mutate; Mutation; Peptide Sequence Determination; Pharmaceutical Preparations; Phase; Physiological; Procedures; Process; Protein Biosynthesis; Proteins; Reading; Reading Frames; Reporter; Research; Research Personnel; Ribosomes; Risk; Scanning; Scientist; Screening procedure; Site; System; Therapeutic Agents; Time; Toxic effect; Translation Initiation; Translations; United States National Institutes of Health; Universities; Variant; Work; Yeasts; chemical synthesis; disease-causing mutation; drug development; in vivo; kidney cell; medical schools; molecular mechanics; novel; novel strategies; preference; small molecule; tool

DESCRIPTION (provided by applicant): Translation initiation is a key point in the regulation of gene expression. A central event in this process is the selection of the initiation codon in the mRNA. If the incorrect initiation codon is selected by the translational machinery, a miscoded protein will be produced, which could have disastrous consequences for the cell. My lab has developed a variety of in vitro tools for studying this key step in protein synthesis, and, together with Alan Hinnebusch's lab at the NIH/NICHD, we have recently made significant strides in understanding the molecular mechanics of start codon recognition. One valuable set of tools we lack, however, are small molecules that can modulate the fidelity of initiation codon recognition by the translational machinery. To address this need, we will conduct a screen for compounds that alter the fidelity of initiation codon recognition in vivo in yeast using a dual luciferase reporter system. The most promising of these compounds will be evaluated for efficacy, toxicity and general mode of action. In addition to their value as research tools, compounds discovered in this screen would be potentially important leads for drug development. Variants of many serious genetic diseases are caused by mutations involving initiation codons, and it is possible that these diseases could be ameliorated by drugs that alter the fidelity of start codon recognition, allowing sufficient protein to be produced from the mRNA with the aberrant start site. Compounds that alter the fidelity of start codon recognition might also prove useful for the development of novel anticancer or antifungal agents. This is a high-risk project and a new direction for my lab. However, if even one effective compound is discovered it would be a major breakthrough for the field of eukaryotic translation research and could provide an important starting point for future drug development. Proteins are the workhorses of our cells. They do the essential tasks that keep cells alive and allow them to grow, divide, and specialize into specific types, such as heart or kidney cells. Proteins are made by a cellular machine called the ribosome that reads information contained in our genes to make the corresponding proteins. A messenger molecule called messenger RNA (mRNA) carries a gene's instructions to the ribosome. For the instructions to make sense, the ribosome needs to start reading the mRNA at the correct starting point. When physiological changes in the cell or mutations cause a change in the starting point, diseases such as phenylketonuria, Tay-Sachs and diabetes insipidus can result. Currently there are no known drugs that can push the ribosome to start at the correct point on altered mRNAs, ameliorating these and other diseases. Using a vast collection of chemical compounds housed and maintained by the National Institutes of Health and Johns Hopkins University School of Medicine, we will search for molecules that alter how the ribosome selects the starting point in the mRNA. These compounds could ultimately be developed into drugs to treat diseases such as Tay-Sachs, as well as certain types of infections or cancer. These chemicals would also be valuable tools for scientists seeking to better understand how the ribosome works. A deeper understanding of this complex cellular machine could lead to new approaches to treat the myriad diseases that affect the ribosome's function.

描述（申请人提供）：翻译起始是基因表达调控的关键点。这个过程的中心事件是mRNA中起始密码子的选择。如果翻译机器选择了错误的起始密码子，就会产生错误编码的蛋白质，这可能对细胞产生灾难性的后果。我的实验室已经开发了各种体外工具来研究蛋白质合成中的这一关键步骤，并且，与NIH/NICHD的Alan Hinnebusch实验室一起，我们最近在理解起始密码子识别的分子机制方面取得了重大进展。然而，我们缺乏一套有价值的工具，那就是可以调节翻译机制对起始密码子识别保真度的小分子。为了解决这一需求，我们将进行筛选的化合物，改变保真度的起始密码子识别在酵母体内使用双荧光素酶报告系统。将对这些化合物中最有前途的化合物的功效、毒性和一般作用模式进行评价。除了作为研究工具的价值外，在这种筛选中发现的化合物将成为药物开发的潜在重要线索。许多严重遗传疾病的变异是由涉及起始密码子的突变引起的，并且这些疾病可能可以通过改变起始密码子识别的保真度的药物来改善，从而允许从具有异常起始位点的mRNA产生足够的蛋白质。改变起始密码子识别保真度的化合物也可能被证明可用于开发新型抗癌剂或抗真菌剂。这是一个高风险的项目，也是我实验室的一个新方向。然而，如果发现一种有效的化合物，这将是真核生物翻译研究领域的重大突破，并可能为未来的药物开发提供重要的起点。蛋白质是我们细胞的主要组成部分。它们完成维持细胞存活的基本任务，并允许它们生长，分裂和专门化为特定类型，如心脏或肾脏细胞。蛋白质是由一种叫做核糖体的细胞机器制造的，核糖体读取我们基因中包含的信息，制造相应的蛋白质。一种称为信使RNA（mRNA）的信使分子将基因的指令传递给核糖体。为了使指令有意义，核糖体需要在正确的起点开始阅读mRNA。当细胞的生理变化或突变导致起点改变时，可能导致苯丙酮尿症、泰-萨克斯病和尿崩症等疾病。目前还没有已知的药物可以推动核糖体在改变的mRNA上的正确位置开始，从而改善这些和其他疾病。利用美国国立卫生研究院和约翰霍普金斯大学医学院收藏和维护的大量化合物，我们将寻找改变核糖体如何选择mRNA起始点的分子。这些化合物最终可能被开发成治疗泰-萨克斯病等疾病以及某些类型的感染或癌症的药物。这些化学物质也将是科学家寻求更好地了解核糖体如何工作的宝贵工具。对这种复杂的细胞机器的更深入了解可能会导致治疗影响核糖体功能的无数疾病的新方法。