Assembly Mechanisms of RNA-Protein Complexes for Genetic Control

非编码RNA形成细胞的蛋白质合成和剪接机制，并在 基因表达的所有阶段。所有的RNA都必须开始折叠并与它们的蛋白质伙伴相互作用 因为它们是被合成的。非编码的表达、组装和本地化不当 RNA与癌症、神经退行性变和发育异常有关。虽然 数以千计的新的非编码RNA已经被发现，我们对它们是如何被发现的知之甚少 实际执行他们的职能。该研究计划的目标是确定以下原则 可以预测RNA-蛋白质复合体大小的RNA-蛋白质组装和动力学 以及小型非编码RNA如何快速找到它们的调控目标。这些 概念将使用两个模型系统解决：核糖体生物合成和后 细菌小RNA和Hfq对转录的调控。核糖体是一个重要的例子。 因为核糖体是在生长的细胞中持续合成的 并受到生长速度和应激反应途径的严格调控。新型单分子 荧光和细胞内RNA结构探测方法将被用来可视化RNA如何 折叠和蛋白质结合与转录结合在一起。小的非编码RNA碱基对 MRNA控制mRNA的表达，是一种普遍存在的基于RNA的调节形式。单人 分子荧光、自然质谱学和细菌报告分析将揭示 Hfq RNA伴侣分子在细菌中回收小的非编码RNA，并在 这些RNA找到了它们合适的目标。这项研究的结果将导致新的概念 与RNA-蛋白质复合体在正常生长期间如何忠实地组装有关，以及为什么某些 RNA很容易受到突变、压力或外来RNA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.