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，内含子是通过大型动态剪接体复合体从前 mRNA 中剪接出来的。已经表明，人类绝大多数遗传性疾病是由内含子的不正确去除引起的。人类中的 SR 蛋白是重要的剪接因子，已被证明对于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续步骤中发挥重要作用。 SR 蛋白水平的改变已被证明会导致许多人类疾病，例如癌症和艾滋病毒。考虑到九种蛋白 SR 家族、组织特异性表达水平以及选择性剪接中的复杂作用，在人类中区分单个 SR 蛋白的作用已被证明是困难的。为了区分 SR 蛋白在遗传易处理系统中全基因组的作用，将利用单细胞模型生物裂殖酵母。粟酒裂殖酵母中只有两种SR蛋白，其许多内含子具有与人类相似的特征。鉴于粟酒裂殖酵母中许多内含子的不同内含子结构，仅测试了少数内含子对 SR 蛋白的最佳剪接的依赖性。此外，许多已证明与人类 SR 蛋白相互作用的剪接因子也存在于粟酒裂殖酵母中，尽管仅观察到了其中的少数相互作用，并且目前缺乏彻底的分析。利用酵母独有的遗传技术以及根据人类实验改编的生化测定，拟议的研究将确定 SR 蛋白如何与粟酒裂殖酵母中的剪接体 (AIM1) 相互作用，以及它们在全基因组范围内有效去除中发挥的作用。该模型生物 (AIM2) 中存在许多独特的内含子。此外，拟议的研究将通过分析特定内含子 (AIM3) 上的剪接体组装来确定 SR 蛋白如何影响粟酒裂殖酵母剪接的机制。对 SR 蛋白如何与粟酒裂殖酵母中不同的内含子组一起发挥作用的基本了解将有助于了解 SR 蛋白的一般机制以及整个剪接体加工前 mRNA 的基本过程。 公共健康相关性：真核生物基因表达的重要步骤之一是通过称为剪接体的大型动态复合体从前 mRNA 中去除内含子。内含子去除错误通常会导致癌症、艾滋病毒、严重的神经退行性疾病脊髓性肌萎缩症 (SMA) 和其他人类疾病。本研究的目的是了解称为 SR 蛋白的重要剪接因子家族如何在最佳去除内含子中发挥作用。