Mechanism of Protein Synthesis and Translational Control

蛋白质合成与翻译控制机制

• 批准号: 10207047
• 负责人: SIMPSON JOSEPH
• 金额: $ 35.46万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN DIEGO
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-06-01 至 2026-05-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10207047
• 关键词: American; Antiviral Agents; Binding; Biochemical; Brain; Cells; Cellular Stress; Cessation of life; Childhood; Code; Complex; Conserved Sequence; Data; Development; Disease; Drug Targeting; Eukaryotic Initiation Factor-4E; FMR1; FXR1 gene; FXR2 gene; Fragile X Syndrome; Gene Silencing; Genes; Goals; In Vitro; Influenza A virus; Intellectual functioning disability; Knowledge; Mammals; Messenger RNA; Molecular; Patients; Pharmaceutical Preparations; Poly(A)-Binding Proteins; Protein Biosynthesis; Proteins; Public Health; RNA-Binding Proteins; Regulation; Research; Resistance; Seizures; System; Techniques; Translation Initiation; Translation Process; Translations; Untranslated Regions; Viral; Virus Diseases; autistic behaviour; biophysical techniques; influenza infection; insight; mRNA Expression; paralogous gene; programs; recruit

Project Summary Our research program focuses on the mechanism of eukaryotic protein synthesis and translational control. The two projects we are currently studying are: (1) the mechanism of translational control by the fragile X mental retardation protein (FMRP), and (2) the mechanism of translation initiation on influenza A virus (IAV) mRNAs. Fragile X syndrome is a disease that afflicts about 100,000 Americans and about 3 million people worldwide, resulting in intellectual disability, childhood seizures, and autistic behavior in the patients. The disease is caused by the transcriptional silencing of the fragile X mental retardation 1 gene (FMR1). FMR1 gene codes for an RNA-binding protein, FMRP, which is highly expressed in the brain and is essential for the normal development of the brain. Mammals have two autosomal paralogs of FMRP designated as fragile X related 1 and 2 (FXR1 and FXR2) proteins. FMRP, FXR1 and FXR2 have been implicated in regulating the translation of several mRNAs. However, the precise mechanism by which these proteins regulate the expression of these mRNAs is unknown. The goal of the first project is to understand the molecular mechanism underlying the regulation of protein synthesis by FMRP, FXR1 and FXR2. We will use a robust in vitro translation system, biochemical techniques and quantitative biophysical methods to significantly advance our understanding of the molecular mechanism used by FMRP, FXR1 and FXR2 to regulate protein synthesis. Results of these studies will provide useful insights in identifying potential drug targets to treat fragile X syndrome. The goal of the second project is to investigate the mechanism of translation initiation by IAV mRNAs. IAV is responsible for several thousand deaths annually and is a severe threat to global public health. We have new data that indicate that IAV mRNAs may use a non-canonical mechanism of translation initiation. Our studies show that poly A binding protein 1 (PABP1) binds to the highly conserved sequences present in the 5’-UTR of IAV mRNAs. Additionally, we show that the translation of the IAV mRNA is more resistant to the inactivation of eukaryotic initiation factor 4E (eIF4E) compared to a control mRNA. We hypothesize that the recruitment of PABP1 to the viral 5’-UTRs tethers eIF4G and promote the assembly of the translation initiation complex in an eIF4E-independent manner. This may favor the translation of IAV mRNAs under cellular stress conditions in the cell, which is known to reduce the activity of eIF4E. We will determine whether the binding of PABP1 to the 5’- UTR of IAV mRNAs is essential for translation initiation and the viral cycle using in vitro techniques and cellular IAV infection studies. Our research will lead to fundamental new knowledge about the process of translation initiation on IAV mRNAs, which could help in the development of new antiviral drugs.

项目概要 我们的研究项目专注于真核蛋白质合成和翻译机制 我们目前正在研究的两个项目是：（1）脆弱的翻译控制机制。 X智力低下蛋白（FMRP），以及（2）甲型流感病毒（IAV）的翻译起始机制 脆性 X 综合征是一种困扰着大约 10 万美国人和大约 300 万人的疾病。 在世界范围内，导致患者出现智力障碍、儿童癫痫发作和自闭症行为。 该疾病是由脆性 X 智力低下 1 基因 (FMR1) 的转录沉默引起的。 编码 RNA 结合蛋白 FMRP，该蛋白在大脑中高度表达，对于正常的生命活动至关重要 哺乳动物有两个 FMRP 常染色体旁系同源物，被指定为脆性 X 相关 1。 FXR1 和 2（FXR1 和 FXR2）蛋白与 FMRP、FXR1 和 FXR2 的翻译调节有关。 然而，这些蛋白质调节这些表达的精确机制。 第一个项目的目标是了解 mRNA 的分子机制。 我们将使用强大的体外翻译系统来调节 FMRP、FXR1 和 FXR2 的蛋白质合成。 生化技术和定量生物物理方法显着增进我们对 FMRP、FXR1 和 FXR2 用于调节蛋白质合成的分子机制。 将为识别治疗脆性 X 综合征的潜在药物靶点提供有用的见解。 第二个项目的目标是研究 IAV mRNA 的翻译起始机制。 IAV 每年造成数千人死亡，对全球公共卫生构成严重威胁。 新数据表明 IAV mRNA 可能使用非规范的翻译起始机制。 显示聚 A 结合蛋白 1 (PABP1) 与 5'-UTR 中存在的高度保守序列结合 此外，我们发现 IAV mRNA 的翻译对 IAV mRNA 的失活具有更强的抵抗力。 与对照 mRNA 相比，我们努力招募真核起始因子 4E (eIF4E)。 PABP1 与病毒 5’-UTR 结合 eIF4G 并促进翻译起始复合物在 eIF4E 独立的方式可能有利于 IAV mRNA 在细胞应激条件下的翻译。 细胞，已知它会降低 eIF4E 的活性，我们将确定 PABP1 是否与 5'- 结合。 使用体外技术和细胞技术，IAV mRNA 的 UTR 对于翻译起始和病毒周期至关重要 我们的研究将带来有关翻译过程的基本新知识。 IAV mRNA 的启动，这可能有助于开发新的抗病毒药物。