Roles of S. pombe SR proteins in spliceosome function and assembly

粟酒裂殖酵母 SR 蛋白在剪接体功能和组装中的作用

• 批准号: 8312142
• 负责人: Michael Charles Marvin
• 金额: $ 4.92万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN FRANCISCO
• 依托单位国家: 美国
• 财政年份: 2012
• 资助国家: 美国
• 起止时间: 2012-04-01 至 2015-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8312142
• 关键词: Affect; Alleles; Alternative Splicing; Animal Model; Architecture; Arginine; Binding; Biochemical; Biological Assay; Characteristics; Complex; Consensus; Defect; Dipeptides; Enhancers; Eukaryota; Excision; Failure; Family; Fission Yeast; Functional RNA; Gel; Gene Expression; Genes; Genetic; Genetic Techniques; Goals; Growth; HIV; Hereditary Disease; Human; Individual; Introns; Knock-out; Lead; Libraries; Malignant Neoplasms; Mammals; Modeling; Neurodegenerative Disorders; Organism; Partner in relationship; Phosphorylation; Play; Positioning Attribute; Process; Protein Splicing; Proteins; Pyrimidine; RNA; RNA Splicing; RNA-Binding Proteins; Reaction; Regulation; Relative (related person); Reliance; Research; Role; Serine; Site; Small Nuclear RNA; Spinal Muscular Atrophy; Spliceosome Assembly Pathway; Spliceosomes; Structure; System; Testing; Tissues; Yeasts; exon skipping; genome-wide; human disease; insight; mRNA Precursor; mutant; protein function; research study

DESCRIPTION (provided by applicant): An essential step in the expression of the majority of genes in humans is the removal of introns, which are spliced out of pre-mRNA by the large and dynamic spliceosome complex. It has been indicated that a significant majority of hereditary disease in humans is caused by the incorrect removal of introns. SR proteins in humans are important splicing factors that have been shown to be essential for spliceosome function and regulation. More specifically, they have been shown to play essential roles in spliceosome assembly as well as later steps of the splicing reaction. Altered levels of SR proteins have been shown to result in many human diseases such as cancer and HIV. Distinguishing the roles of individual SR proteins has proven difficult in humans given the nine-protein SR family, tissue specific expression levels, and complex roles in alternative splicing. In order to distinguish the roles of SR proteins genome-wide in a genetically tractable system, the unicellular model organism Schizosaccharomyces pombe will be utilized. There are only two SR proteins in S. pombe with many introns containing similar characteristics with humans. Given the diverse intron structure of the many introns in S. pombe only a few introns have been tested for reliance on SR proteins for optimal splicing. In addition, many of the splicing factors that have been shown to interact with human SR proteins are also present in S. pombe although only a few of these interactions have been observed and a thorough analysis is currently lacking. Using genetic techniques that are uniquely available to yeast as well as biochemical assays adapted from experiments in humans, proposed studies will determine how SR proteins interact with the spliceosome in S. pombe (AIM1) and what role they play genome-wide in the efficient removal of the many unique introns that are present in this model organism (AIM2). In addition, proposed studies will determine the mechanism of how SR proteins influence splicing in S. pombe by analyzing spliceosome assembly on specific introns (AIM3). The fundamental understanding of how SR proteins function with the diverse set of introns in S. pombe will inform general mechanisms of SR proteins and the essential process of pre-mRNA processing by the spliceosome as a whole. PUBLIC HEALTH RELEVANCE: One of the essential steps of gene expression in eukaryotes is the removal of introns by the large and dynamic complex known as the spliceosome from pre-mRNA. Errors in intron removal often can result in cancer, HIV, the severe neurodegenerative disorder spinal muscular atrophy (SMA) and other human diseases. The goal of the proposed research is to understand how the important family of splicing factors called SR proteins function in the optimal removal of introns.

描述（由申请人提供）：人类大多数基因表达的一个重要步骤是去除内含子，内含子是由大而动态的剪接体复合物从Pre-mRNA中剪接出来的。已经表明，人类中大部分遗传性疾病的大部分是由于内含子的错误去除引起的。人类中的SR蛋白是重要的剪接因子，已证明对剪接体功能和调节至关重要。更具体地说，它们已被证明在剪接体组装以及剪接反应的后期步骤中起着重要作用。 SR蛋白水平的改变已被证明会导致许多人类疾病，例如癌症和HIV。鉴于9蛋白SR家族，组织特异性表达水平以及在替代剪接中的复杂作用，从人类中区分各个SR蛋白的作用已被证明很难。为了区分SR蛋白质全基因组在遗传障碍系统中的作用，将利用单细胞模型生物体菌chizosaccharomyces pombe。 S. pombe中只有两个SR蛋白，其中包含与人类相似特征的许多内含子。鉴于S. pombe中许多内含子的内含子结构的不同，只有少数内含子经过了依赖SR蛋白的最佳剪接测试。此外，尽管仅观察到其中一些相互作用，并且目前缺乏彻底的分析，但已显示出与人类SR蛋白相互作用的许多剪接因子也存在。拟议的研究使用适用于人类实验的遗传技术以及适合于人类实验的生化测定方法，将决定SR蛋白如何与S. Pombe中的剪接体相互作用（AIM1）（AIM1）以及它们在有效地删除该模型生物体中存在的许多独特内含子（AIM AIMISISMISSISMISISISMISSISMISSISMISSISMISSISMISSISM）中扮演的基因组的作用。此外，提出的研究将通过分析特定内含子的剪接组装来确定SR蛋白如何影响S. Pombe剪接的机制（AIM3）。对SR蛋白在S. Pombe中的多种内含子的发挥作用的基本理解将为SR蛋白的一般机制提供信息，以及整个剪接体通过剪接体进行前MRNA加工的基本过程。 公共卫生相关性：真核生物中基因表达的重要步骤之一是从pre-mRNA中被称为剪接体的大型且动态的复合物去除内含子。内含子去除的错误通常会导致癌症，HIV，严重的神经退行性疾病脊柱肌肉萎缩（SMA）和其他人类疾病。拟议的研究的目的是了解称为SR蛋白的重要剪接因子系列如何在最佳去除内含子中起作用。