A genomic approach to studying the life cycle of intron lariats

研究内含子套索生命周期的基因组方法

• 批准号: 9043905
• 负责人: William G Fairbrother
• 金额: $ 41.42万
• 依托单位: BROWN UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-07-16 至 2018-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9043905
• 关键词: 3' Splice Site; Affect; Area Analyses; Binding; Biochemical; Biological; Biological Assay; Biology; Catalysis; Cells; Computer software; DNA Polymerase II; Data; Defect; Degradation Pathway; Dependency; Detection; Disease; Elderly; Enzymes; Event; Exons; Exonuclease; Genes; Genetic Transcription; Genome; Genomic approach; Goals; Grant; Human; Human Genome; In Vitro; Indium; Introns; Kinetics; Knowledge; Lead; Life; Life Cycle Stages; Location; Maps; Medical Genetics; Messenger RNA; Methods; MicroRNAs; Modeling; Mus; Mutation; Nuclear; Nucleotides; Pathway interactions; Process; Product Recycling; Publishing; RNA; RNA Splicing; RNA-Binding Proteins; Reading; Recycling; Repression; Research; Role; Sampling; Signal Transduction; Site; Small Nucleolar RNA; Spliceosomes; System; Testing; Time; Tissues; Transcript; Untranslated RNA; Upstream Enhancer; Variant; Yeasts; clinical sequencing; deep sequencing; exosome; in vivo; insight; mRNA Precursor; novel; public health relevance; research study; scale up; success; transcription factor

DESCRIPTION (provided by applicant): Pre-mRNA splicing is a critical and regulated processing event where introns are precisely excised from nascent RNA transcripts. As many as one third of all heritable disease mutations result in splicing defects. This research studies the role of branchpoints in determining splice site selection (3'ss) is utilized in vivo and also the effect branchpoints have on the life cycle o the intron. Each pre-mRNA splicing event creates a lariat and spliced exon junction. While a great deal is known about splice exon junctions almost nothing is known about lariats. By mapping all branchpoints in the human genome, we are opening up a whole new area of analysis. The identification of branchpoints by transcript data will facilitate the interpretation f clinical sequencing data. In addition to the intrinsic value of this data, the successful completio of this proposal will test some hypothesis about the fundamental catalysis and recognition that occurs in vivo in the processing of eukaryotic genes. Studying these intermediates at a system wide level will bring a biochemical-level understanding to hundreds of thousands of processing events. Furthermore, each intron lariat has a lifecycle - created by splicing of a transcribed product, recycled by debranching and degradation. The recycling of introns is vital to replenishing the intracellular levels of free nucleotides and to return splicing factors into activ spliceosomes. Some introns have a second life after splicing as non-coding RNAs (ncRNAs). As we are sampling steady state levels of introns we gain insight into both these processes. Our preliminary data indicates a novel degradation that is devoted to recycling large introns. This proposal seeks to follow this lead by defining the complete degradation pathways and exploring some of the reasons why certain introns appear stabilized.

描述(由申请人提供)： 前mRNA剪接是一个关键的和受调控的加工事件，其中内含子被精确地从新生的RNA转录本中切除。在所有可遗传疾病突变中，多达三分之一会导致剪接缺陷。本研究研究了分支点在决定体内剪接位点选择(3‘s)中的作用以及分支点对内含子生命周期的影响。每个前mRNA剪接事件都会产生一个套索和剪接的外显子连接。虽然人们对剪接外显子连接的了解很多，但几乎没有人知道连接基因。通过绘制人类基因组的所有分支点，我们正在开辟一个全新的分析领域。通过转录本数据识别分支点将有助于临床测序数据的解释。除了这一数据的内在价值之外，这一提议的成功完成还将检验关于真核基因处理过程中体内发生的基本催化和识别的某些假设。在系统范围的水平上研究这些中间体将为数十万过程事件带来生化水平的理解。此外，每个内含子套索都有一个生命周期--通过转录产物的剪接创建，通过去分支和降解进行循环。内含子的循环对于补充细胞内游离核苷酸水平和将剪接因子返回活性剪接体是至关重要的。一些内含子在作为非编码RNA(NcRNAs)剪接后有第二次生命。当我们采样内含子的稳态水平时，我们对这两个过程都有了深入的了解。我们的初步数据表明，一种致力于回收大内含子的新型降解。这项提议试图通过定义完整的降解途径并探索某些内含子似乎稳定的一些原因来追随这一线索。