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Complexity and function of messenger ribonucleoprotein particles

信使核糖核蛋白颗粒的复杂性和功能

基本信息

批准号：
197040498
负责人：
Professorin Dr. Christine Elizabeth Clayton
金额：
--
依托单位：
Zentrum für Molekulare Biologie (ZMBH)
依托单位国家：
德国
项目类别：
Research Grants
财政年份：
2011
资助国家：
德国
起止时间：
2010-12-31 至 2014-12-31
项目状态：
已结题

来源：
https://gepris.dfg.de/gepris/projekt/197040498?language=en
关键词：
Complexity function messenger ribonucleoprotein particles

项目摘要

Cells determine the final protein output of their genetic program not only by controlling transcription, but also by regulating the localization, stabilities and translation rates of their mRNAs. Our laboratories are investigating mechanisms that control gene expression at the post-transcriptional level in two widely separated branches of eukaryotic evolution: trypanosomes and mammals. Ultimately, the fate of any given mRNA is determined by the ensemble of all associated RNA-binding proteins (RBPs), i.e. the messenger ribonucleoprotein particle (mRNP). Surprisingly little, however, is known about the composition of mRNPs. In this grant application, we want to characterize mRNPs from trypanosomes and mammalian cell lines by purification of polypeptide nascent chains, and by introducing RNA affinity tags into reporter mRNAs. Our specific goals are 1) to characterize the mRNP encoding the Variable Surface Glycoprotein, the most abundant protein of bloodstream trypanosomes; 2) to compare the trypanosome and mammalian mRNPs producing α-tubulin, a highly abundant and conserved protein throughout eukaryotic evolution; and 3) to identify RBPs associated with a novel RNA decay element that we found in the mRNAs encoding mouse and human tumor necrosis factor-α. These studies will not only improve our basic understanding of eukaryotic mRNPs, but also provide methods that are useful in addressing many questions in the field of RNA biology.

细胞决定其遗传程序的最终蛋白质输出不仅通过控制转录，而且通过调节其mRNA的定位，稳定性和翻译速率。我们的实验室正在研究在真核生物进化的两个广泛分离的分支中在转录后水平控制基因表达的机制：锥虫和哺乳动物。最终，任何给定mRNA的命运由所有相关RNA结合蛋白（RBP）的集合体决定，即信使核糖核蛋白颗粒（mRNP）。然而，令人惊讶的是，对mRNP的组成知之甚少。在这项授权申请中，我们希望通过纯化多肽新生链，并通过将RNA亲和标签引入报告基因mRNA来表征锥虫和哺乳动物细胞系中的mRNP。我们的具体目标是：1）表征编码可变表面糖蛋白（血流锥虫中最丰富的蛋白质）的mRNP; 2）比较锥虫和哺乳动物产生α-微管蛋白（一种在真核生物进化过程中高度丰富和保守的蛋白质）的mRNP; 3）鉴定与我们在编码小鼠和人肿瘤坏死因子-α的mRNA中发现的新型RNA衰变元件相关的RBP。这些研究不仅将提高我们对真核mRNP的基本理解，而且还提供了解决RNA生物学领域许多问题的有用方法。