Biogenesis of Small RNAs

描述（由申请人提供）：非编码RNA，如剪接体U snRNA、转移RNA、核糖体RNA和信号识别颗粒和端粒酶的RNA组分，在真核细胞代谢中起关键作用。虽然对这些RNA的功能了解很多，但对这些RNA如何在细胞内折叠成复杂结构以及细胞如何处理错误折叠和有缺陷的RNA知之甚少。我们的长期目标是了解生产性RNA折叠的细胞要求，并揭示细胞检测和降解缺陷RNA的机制。我们的具体策略是专注于与新生RNA相互作用的蛋白质，在酵母酿酒酵母中使用遗传学和生物化学。我们以前的工作表明，一个保守的蛋白质，La蛋白，保护新生的非编码RNA从核酸酶，协助前tRNA折叠和接口与缺陷RNA的监视途径。我们还描述了一个复杂的Sm样蛋白质的功能在核仁RNA的一个子集的生物发生或功能。我们现在的目标是剖析La和其他蛋白质协助非编码RNA生物合成和质量控制的分子机制。我们的第一个目标是使用生物化学，遗传学和结构生物学来确定La如何稳定正确折叠的RNA。我们的第二个目标是阐明La在非编码RNA监视中的作用。我们的第三个目标是描述几个增加不稳定非编码RNA丰度的突变。突变基因的克隆及其产物的表征应该揭示非编码RNA质量控制的新参与者。我们的第四个目标是纯化Sm样蛋白复合物，并确定其在酵母核仁中的作用。由于大部分基因表达是通过非编码RNA介导的，因此对RNA生物发生和质量控制的理解对于揭示细胞逃避正常生长控制的机制和设计新的治疗方法具有广泛的重要性。敷设总结：虽然非编码RNA对细胞功能至关重要，但人们对这些RNA是如何产生的以及细胞如何检测和去除含有错误的RNA知之甚少。通过了解这些过程，有可能设计出治疗非编码RNA功能不正常引起的疾病的方法。

DESCRIPTION (provided by applicant): Noncoding RNAs, such as the spliceosomal U snRNAs, transfer RNAs, ribosomal RNAs and the RNA components of signal recognition particle and telomerase, play critical roles in eukaryotic cell metabolism. Although much is known about the functions of these RNAs, little is known as to how these RNAs fold into complex structures within cells and how cells handle misfolded and defective RNAs. Our long term objective is to understand the cellular requirements for productive RNA folding and to uncover the mechanisms by which cells detect and degrade defective RNAs. Our specific strategy is to focus on proteins that interact with nascent RNAs, using genetics and biochemistry in the yeast Saccharomyces cerevisiae. Our previous work revealed that a conserved protein, the La protein, protects nascent noncoding RNAs from nucleases, assists pre-tRNA folding and interfaces with a surveillance pathway for defective RNAs. We also described a complex of Sm-like proteins that functions in the biogenesis or function of a subset of nucleolar RNAs. Our goal now is to dissect the molecular mechanisms by which La and other proteins assist noncoding RNA biogenesis and quality control. Our first aim is to use biochemistry, genetics and structural biology to determine how La stabilizes correctly folded RNAs. Our second aim is to elucidate the role of La in noncoding RNA surveillance. Our third aim is to characterize several mutations that increase the abundance of unstable noncoding RNAs. Cloning of the mutant genes and characterization of their products should reveal new players in noncoding RNA quality control. Our fourth aim is to purify the Sm-like protein complex and determine its role in the yeast nucleolus. As much of gene expression is mediated through noncoding RNAs, an understanding of RNA biogenesis and quality control should be of broad importance for uncovering mechanisms by which cells escape normal growth controls and for designing new therapeutics. Lay summary: Although noncoding RNAs are critical for cell function, little is known as to how these RNAs are made and how cells detect and remove RNAs that contain errors. By understanding these processes, it may be possible to design treatments for diseases caused by noncoding RNAs that do not function properly.