Quantitative Analysis of RNA Folding in Yeast

酵母中 RNA 折叠的定量分析

• 批准号: 6938308
• 负责人: JASON HARGER
• 金额: $ 3.64万
• 依托单位: SCRIPPS RESEARCH INSTITUTE, THE
• 依托单位国家: 美国
• 财政年份: 2005
• 资助国家: 美国
• 起止时间: 2005-04-01 至 2006-01-17
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6938308
• 关键词: RNA; catalyst; chemical cleavage; chemical structure function; conformation; enzyme activity; molecular assembly /self assembly; nucleic acid structure; postdoctoral investigator; ribozymes; yeasts

DESCRIPTION (provided by applicant): The dynamics and equilibria of RNA conformational changes are critical features of the multitude of RNA-mediated reactions that take place in living cells. RNA enzymes or ribozymes have provided the gold standard for RNA structure-function analyses for the past two decades because they reveal their assembly into functional structures through catalytic activity. But, despite two decades of research our understanding of the basic mechanisms of RNA assembly remains limited. The hairpin ribozyme, encoded in the (-)TRSV satellite RNA, is a small catalytic motif that catalyzes sequence-specific reversible self-cleavage. Existing structural, biochemical, and kinetic data for the hairpin ribozyme, and the availability of a yeast-based in vivo reporter system make this motif and ideal model for studying intracellular RNA assembly pathways. This system will be used to determine the kinetic and thermodynamic influences on ribozyme assembly during transcription and translation inside living cells, and to monitor whether RNAs interact within discrete folding domains during co-transcriptional ribozyme assembly. Ultimately this work will improve our understanding of the basic principles governing RNA assembly in vivo.

描述(由申请人提供)：RNA构象变化的动态和平衡是在活细胞中发生的大量RNA介导的反应的关键特征。在过去的二十年里，RNA酶或核酶为RNA结构-功能分析提供了黄金标准，因为它们揭示了它们通过催化活性组装成功能结构。但是，尽管经过了20年的研究，我们对RNA组装的基本机制的了解仍然有限。发夹状核酶编码在(-)TRSV卫星RNA中，是一个小的催化基序，催化序列特异性的可逆自切割。现有的发夹状核酶的结构、生化和动力学数据，以及基于酵母的体内报告系统的可用性，使这一基序成为研究细胞内RNA组装途径的理想模型。该系统将被用来确定在活细胞内转录和翻译过程中对核酶组装的动力学和热力学影响，以及在共转录核酶组装过程中RNA是否在离散的折叠结构域内相互作用。最终，这项工作将提高我们对体内RNA组装的基本原理的理解。