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.

项目摘要 我们的研究计划着重于真核蛋白质合成和翻译的机制 控制。我们目前正在研究的两个项目是：（1）脆弱的转化控制机制 X智力低下蛋白（FMRP），以及（2）影响Za病毒（IAV）的翻译启动机理 mrnas。脆弱的X综合征是一种疾病，折磨约100,000美国人和约300万人 在全球范围内，导致患者的智力残疾，儿童癫痫发作和加速行为。 疾病是由脆弱X智力低下1基因（FMR1）的转录沉默引起的。 FMR1基因 RNA结合蛋白FMRP的代码，该蛋白在大脑中高度表达，对于正常 大脑的发展。哺乳动物有两个指定为脆弱x相关的FMRP的常染色体旁系同源物1 和2（FXR1和FXR2）蛋白质。 FMRP，FXR1和FXR2在调节翻译时隐含 几个mRNA。但是，这些蛋白质调节这些蛋白的表达的确切机制 mRNA是未知的。第一个项目的目的是了解该分子机制 FMRP，FXR1和FXR2对蛋白质合成的调节。我们将使用强大的体外翻译系统， 生化技术和定量生物物理方法，以显着促进我们对 FMRP，FXR1和FXR2使用的分子机制来调节蛋白质合成。这些研究的结果 将提供有用的见解，以确定治疗脆弱X综合征的潜在药物靶标。 第二个项目的目的是研究IAV mRNA的翻译计划机制。 IAV每年造成数千人死亡，对全球公共卫生构成严重威胁。我们有 新数据表明IAV mRNA可能会使用非典型的翻译起始机制。我们的研究 表明poly A结合蛋白1（PABP1）与存在于5'-utr中的高度组成序列结合 IAV mRNA。此外，我们表明IAV mRNA的翻译对失活的抗性更具 与对照mRNA相比，真核启动因子4E（EIF4E）。我们假设招募 PABP1到病毒5'-UTRS TETHERS EIF4G并促进翻译起始复合物的组装 EIF4E独立的方式。这可能有利于在细胞应力条件下IAV mRNA的翻译 细胞，已知可以减少EIF4E的活性。我们将确定PABP1是否与5'-的结合 IAV mRNA的UTR对于使用体外技术和细胞的翻译开始和病毒循环至关重要 IAV感染研究。我们的研究将导致有关翻译过程的基本新知识 对IAV mRNA的启动，可以帮助开发新的抗病毒药。