Analysis of FGF2 5UTR mRNPs in the control of alternative translation initiation site choice

FGF2 5UTR mRNP 控制替代翻译起始位点选择的分析

• 批准号: 241756384
• 负责人: Privatdozent Dr. Jan Medenbach
• 金额: --
• 依托单位: Lehrstuhl für Biochemie I
• 依托单位国家: 德国
• 项目类别: Research Grants
• 财政年份: 2013
• 资助国家: 德国
• 起止时间: 2012-12-31 至 2014-12-31
• 项目状态: 已结题
• 来源: https://gepris.dfg.de/gepris/projekt/241756384?language=en
• 关键词: Analysis; FGF2; 5UTR; mRNPs; control

The regulation of translation presents an important and critical layer of gene expression control in eukaryotic organisms allowing for spatio-temporal regulation of protein synthesis. It can affect both the amount of protein and the isoform synthesized (quantitative vs. qualitative control of translation). While we are slowly beginning to unravel the principles underlying quantitative regulation of translation, qualitative control still remains mostly enigmatic.Already in the 1980ies it was discovered that alternative translation initiation was employed by a number of mRNAs to produce different protein isoforms from a single RNA species (such as e.g. FGF2, C-MYC, VEGF). Certain cellular conditions can trigger the translation of N-terminally extended protein isoforms which exhibit markedly different activities compared to their smaller counterparts. Exemplarily, in contrast to the small protein isoform, high molecular weight (HMW) fibroblast growth factor 2 (FGF2) proteins are not secreted from the cell but translocate into the nucleus where they exhibit immortalizing activities. Despite the clinical importance of the various FGF2 peptides it remains unknown how production of the individual protein isoforms is controlled. Recent technological advances now allow us to examine ribonucleoproteins (RNPs) in a highly parallel fashion, probing simultaneously for a multitude of protein-RNA interactions. By further developing, adapting and applying these high throughput techniques to FGF2 mRNA (and a panel of additional RNAs of interest) I plan to interrogate RNP dynamics under different cellular conditions in unprecedented detail. Combining these high throughput approaches with reductionist, mechanistic experimentation will allow novel insights into the regulation of translation initiation and start codon choice on FGF2 mRNA, a truly important but yet poorly understood process in biology.

翻译调节是真核生物中基因表达控制的一个重要且关键的层，允许蛋白质合成的时空调节。它可以影响蛋白质的数量和合成的亚型（翻译的定量与定性控制）。虽然我们正在慢慢地开始解开翻译定量调控的原理，但定性控制仍然是一个谜。早在 20 世纪 80 年代，人们就发现许多 mRNA 采用替代翻译起始，从单一 RNA 物种（例如 FGF2、C-MYC、VEGF）中产生不同的蛋白质亚型。某些细胞条件可以触发 N 末端延伸的蛋白质亚型的翻译，与较小的对应物相比，其表现出明显不同的活性。示例性地，与小蛋白亚型相反，高分子量（HMW）成纤维细胞生长因子2（FGF2）蛋白不从细胞分泌，而是易位到细胞核中，在那里它们表现出永生化活性。尽管各种 FGF2 肽具有临床重要性，但如何控制各个蛋白质亚型的产生仍然未知。最近的技术进步现在使我们能够以高度并行的方式检查核糖核蛋白 (RNP)，同时探测多种蛋白质-RNA 相互作用。通过进一步开发、调整和应用这些高通量技术到 FGF2 mRNA（以及一组其他感兴趣的 RNA），我计划以前所未有的细节探究不同细胞条件下的 RNP 动态。将这些高通量方法与还原论、机械实验相结合，将为 FGF2 mRNA 的翻译起始调节和起始密码子选择提供新的见解，这是生物学中一个真正重要但仍知之甚少的过程。