Assembly Mechanisms of RNA-Protein Complexes for Genetic Control

非编码RNA形成了细胞的蛋白质合成和剪接机制，并在蛋白质合成过程中起作用。 基因表达的各个阶段。所有RNA都必须开始折叠并与其蛋白质伴侣相互作用 因为它们正在被合成。非编码的表达、组装和定位不当 RNA与癌症、神经变性和发育异常有关。虽然 尽管已经发现了数千种新的非编码RNA，但我们对它们是如何被发现的知之甚少。 物理地执行其功能。这项研究计划的目标是确定原则， RNA-蛋白质组装和动力学可以预测RNA-蛋白质复合物的大小 以及小的非编码RNA如何快速找到它们的调控靶点。这些 概念将解决使用两个模型系统：核糖体生物合成和后， 通过细菌小RNA和Hfq的转录调控。核糖体是一个重要的例子 因为核糖体在生长的细胞中不断合成， 并受到生长速度和应激反应途径的严格调控。新单分子 荧光和细胞内RNA结构探测方法将用于可视化RNA如何 折叠和蛋白质结合与转录偶联。小的非编码RNA碱基对， mRNA调控mRNA的表达，是一种普遍存在的基于RNA的调控形式。单个 分子荧光，天然质谱和细菌报告分析将揭示如何 Hfq RNA伴侣蛋白在细菌中降解小的非编码RNA，并在细菌中增加速率。 让这些RNA找到合适的目标这项研究的结果将导致新的概念 与RNA-蛋白质复合物在正常生长过程中如何忠实地组装以及为什么某些 RNA易受突变、压力或来自外源RNA的竞争的影响。

Non-coding RNA forms the cell’s machineries for protein synthesis and splicing, and acts during all stages of gene expression. All RNAs must begin to fold and interact with their protein partners as they are being synthesized. Improper expression, assembly and localization of non-coding RNA has been linked to cancer, neurodegeneration and developmental abnormalities. Although thousands of new non-coding RNAs have been discovered, we know little about how they physically carry out their functions. The goal of this research program is to define principles of RNA-protein assembly and dynamics that can predict how large RNA-protein complexes are formed and how small non-coding RNAs rapidly search out their regulatory targets. These concepts will be addressed using two model systems: ribosome biosynthesis and post- transcriptional regulation by bacterial small RNA and Hfq. Ribosomes are an important example of co-transcriptional assembly because ribosomes are continually synthesized in growing cells and tightly regulated by growth rate and by stress response pathways. New single molecule fluorescence and in-cell RNA structure probing approaches will be used to visualize how RNA folding and protein binding is coupled to transcription. Small non-coding RNAs base pair with mRNA to control mRNA expression, and are a ubiquitous form of RNA-based regulation. Single molecule fluorescence, native mass spectrometry and bacterial reporter assays will reveal how the Hfq RNA chaperone recycles small non-coding RNA in bacteria, and increases the rate at which these RNAs find their proper targets. The results of this research will lead to new concepts related to how RNA-protein complexes assemble faithfully during normal growth and why certain RNAs are vulnerable to mutation, stress, or competition from foreign RNA.