KINETIC/THERMODYNAMIC/STRUCTURAL STUDIES OF RNA FOLDING

描述：RNA的生物学功能与它的 三维结构。 RNA序列如何指定其三级结构 结构是RNA结构生物学的最终目标。 为了 了解RNA折叠是如何完成的，阐明RNA折叠的机制是至关重要的。 RNA结构的热力学稳定性和动力学折叠 通路 本提案的长期目标是确定 热力学和折叠动力学的两个高度结构化的RNA。 在 前两个目标，来自细菌RNase P的核酶组分将是 研究了包含多个结构域的大RNA的折叠。 两个已知的热力学/动力学的紧密性和三级结构 将对中间体进行评价。 域-域交互作用的作用 在折叠将被检查。 折叠也将研究期间 在模拟细胞条件下转录，以确定 RNA可以在体内折叠。 此外，将分离RNase P RNA变体， 其折叠途径可以从野生型核酶的折叠途径改变。 在第三个目标中，将转移RNA作为模型系统进行研究， 理解一个单一的、孤立的三级结构的折叠。 的tRNA 研究不仅会阐明tRNA如何折叠，更重要的是， 在物理化学分析中产生一个概念框架， 所有的RNA 这一建议的结果将加深我们对以下问题的理解： RNA如何折叠成特定的三级结构， RNA折叠的基本原理

DESCRIPTION: The biological function of RNA is intimately related to its three-dimensional structure. How an RNA sequence specifies its tertiary structure is an ultimate goal of RNA structural biology. In order to understand how RNA folding is accomplished, it is essential to elucidate the thermodynamic stability of the RNA structure and the kinetic folding pathway. The long-term objective of this proposal is to determine the thermodynamics and the folding kinetics of two highly structured RNAs. In the first two aims, the ribozyme component from bacterial RNase P will be studied to understand folding of a large RNA containing multiple domains. The compactness and tertiary structure of two known thermodynamic/kinetic intermediates will be evaluated. The role of the domain-domain interactions in folding will be examined. Folding will also be studied during transcription under simulated cellular conditions to determine how large RNAs may fold in vivo. In addition, RNase P RNA variants will be isolated, whose folding pathway may be altered from that of the wild-type ribozyme. In the third aim, transfer RNA will be studied as a model system to understand folding of a single, isolated tertiary structure. The tRNA studies will not only elucidate how tRNAs fold, but also more importantly, generate a conceptual framework in physico-chemical analysis for folding of all RNAs. The results from this proposal will deepen our understanding in how an RNA folds into a particular tertiary structure and establish the fundamental principles of RNA folding.