RNA-based mechanisms in nuclear steps of gene expression

基因表达核步骤中基于 RNA 的机制

基本信息

批准号：
10673582
负责人：
DAVID A BROW
金额：
$ 31.99万
依托单位：
UNIVERSITY OF WISCONSIN-MADISON
依托单位国家：
美国
项目类别：
财政年份：
2016
资助国家：
美国
起止时间：
2016-07-01 至 2026-07-31
项目状态：
未结题

项目摘要

Project Summary RNA-RNA and RNA-protein interactions lie at the heart of essential steps of the eukaryotic gene expression pathway. Defects in these interactions due to inherited mutations can result in age- dependent degeneration of the retina, motor neurons, and other neural tissues. The proposed studies will result in a better understanding of RNA-based mechanisms of gene expression in both the normal and disease states. In the next five years we will pursue three main goals, using the yeast Saccharomyces cerevisiae as a facile model system. First, we will examine the molecular mechanism of activation of the spliceosome for the first catalytic step of pre-mRNA splicing. This process requires allosteric signal transmission through RNA and protein over distances of 100 angstroms or more and results in large-scale remodeling of the spliceosome to allow progression of the splicing cycle. Second, we will elucidate the basis for regulation of expression of a key enzyme in purine nucleotide metabolism, IMPDH, interrogating both transcriptional and post-transcriptional steps. Inherited alterations in a regulatory domain of IMPDH and in proteins that direct spliceosome activation are associated with autosomal dominant retinitis pigmentosa, which leads to progressive blindness. We will look for commonalities between the two processes that might explain the highly specific pathological consequences of these disease mutations. Third, we will examine the functions of the hnRNP protein Hrp1 in the regulation of elongation and termination by RNA polymerase II. Hrp1 is structurally related to human hnRNP proteins in which inherited substitutions cause neurodegenerative disorders, including ALS and FTLD. Furthermore, Hrp1 exhibits a similar propensity to form intracellular aggregates, which are associated with pathology of the human proteins. A more complete understanding of these fundamental processes should lead to more accurate diagnosis and prognosis of diseases caused by alterations in nuclear RNA-binding proteins, and may ultimately result in new therapeutic approaches. Furthermore, the proposed studies will illuminate basic mechanisms of eukaryotic gene expression that can be exploited for synthetic biology and biotechnology.

项目摘要 RNA-RNA和RNA-蛋白质相互作用位于真核基因基本步骤的核心表达途径。由于遗传突变引起的这些相互作用的缺陷可能导致年龄视网膜，运动神经元和其他神经组织的依赖性变性。提议研究将使人们更好地了解基于RNA的基因表达机制正常和疾病状态。在接下来的五年中，我们将实现三个主要目标酿酒酵母作为一种简便的模型系统。首先，我们将检查剪接体激活的分子机制，用于前MRNA的第一个催化步骤剪接。此过程需要通过RNA和蛋白质的变构信号传递 100埃或更多的距离，并导致剪接体大规模重塑至允许剪接周期的进展。其次，我们将阐明监管的基础在嘌呤核苷酸代谢中的键酶的表达，IMPDH，询问两者转录和转录后步骤。在监管领域的继承变化 IMPDH和直接剪接体激活的蛋白质与常染色体有关显性视网膜炎色素炎，导致渐进的失明。我们会寻找这两个过程之间的共同点可能解释了高度特定的病理这些疾病突变的后果。第三，我们将检查HNRNP的功能 RNA聚合酶II的延长和终止的蛋白HRP1。 HRP1是在结构上与人类HNRNP蛋白相关，其中继承的取代导致神经退行性疾病，包括ALS和FTLD。此外，HRP1表现出类似的形成与人体病理有关的细胞内聚集体的倾向蛋白质。对这些基本过程的更完整的理解应导致更多核RNA结合改变引起的疾病的准确诊断和预后蛋白质，并最终可能导致新的治疗方法。此外，提议研究将阐明可利用的真核基因表达的基本机制合成生物学和生物技术。