Biosynthesis of RNAs

RNA的生物合成

• 批准号: 7033075
• 负责人: CHRISTINE GUTHRIE
• 金额: $ 74.06万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN FRANCISCO
• 依托单位国家: 美国
• 财政年份: 1977
• 资助国家: 美国
• 起止时间: 1977-02-01 至 2008-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7033075
• 关键词: RNA binding protein; RNA biosynthesis; RNA splicing; Saccharomyces cerevisiae; active sites; adenosine triphosphate; conformation; fungal genetics; gene environment interaction; gene expression; gene interaction; genetic regulation; high throughput technology; intracellular transport; messenger RNA; microarray technology; nucleic acid sequence; polymerase chain reaction; ribonucleoproteins; site directed mutagenesis; small nuclear ribonucleoproteins; spliceosomes

DESCRIPTION (provided by applicant): Our long-term interest is in the mechanisms that control the specificity and fidelity of mRNA splicing and export. We study these problems in the budding yeast Saccharomyces cerevisiae in order to exploit its enormous genetic potential. Moreover, our work over the last 20 years has been instrumental in validating the principle - for mRNA splicing - that the basic steps in gene expression are remarkably conserved from yeast to mammals. In the last funding period, we have focused in particular on the role of ATP-driven rearrangements promoted by the DEAD-box family of proteins. A unifying hypothesis that has emerged from our studies is that mRNPs are remodelled extensively via the exchange of RNA:RNA or RNA:protein pairing partners; in some cases, these pairing partners are mutually exclusive. Future experiments will test the notion that these NTP-dependent exchanges are not only physically and temporally correlated, but are functionally interdependent, i.e. coupled. In addition to explicitly addressing the dynamic design principle of the spliceosome's catalytic core, these studies will suggest testable mechanisms for the integration of the major steps of gene expression with one another. Finally, we will exploit our recent development of highthroughput microarray technology to assess the genome-wide impact of specific mutations and environmental stresses on the full set of 250 intron-containing genes. Despite the widely-held notion that splicing in budding yeast is not significantly regulated, our initial analyses suggest that splicing in this single-cell organism is in fact subject to complex combinatorial control. Using a blend of biochemical, genetic, and cell biological approaches, we propose three Specific Aims: 1. To Test a Comprehensive Model for Catalysis and Fidelity at the Second Step of Splicing 2. To Identify Coupled Steps in the Sequential Remodelling of mRNPs that Confer Export Competence 3. To Identify Regulated Pathways in RNA Processing Using Genome-wide Analysis

描述（由申请人提供）：我们的长期利益是在控制mRNA剪接和出口的特异性和保真度的机制中。 我们研究了酿酒酵母的萌芽酵母菌中的这些问题，以利用其巨大的遗传潜力。 此外，过去20年中，我们的工作一直在验证基因表达的基本步骤从酵母到哺乳动物中明显保守的原理（对于mRNA剪接）中发挥了作用。 在最后的资金期间，我们特别关注了由Dead-box蛋白质家族促进的ATP驱动重排的作用。 我们的研究中出现的一个统一的假设是，通过RNA的交换：RNA或RNA：蛋白质配对伴侣；在某些情况下，这些配对伙伴是互斥的。 未来的实验将测试这些依赖NTP的交换不仅在身体和时间上相关的概念，而且在功能上相互依存，即耦合。除了明确解决剪接体的催化核心的动态设计原理外，这些研究还将提出可测试的机制，以将基因表达的主要步骤相互融为一体。最后，我们将利用我们最近的高通量微阵列技术的发展来评估特定突变和环境应力对全套含250个内含子内含器基因的影响的影响。尽管广泛持有的观念认为在萌芽酵母中的剪接并未受到显着调节，但我们的初步分析表明，这种单细胞生物中的剪接实际上受到复杂的组合控制。使用生化，遗传和细胞生物学方法的混合，我们提出了三个具体目标：1。在剪接2的第二步中测试一个综合模型的催化和忠诚度模型。以确定MRNP的顺序重塑中的耦合步骤，以使用RNA处理的调节途径3。