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剪接是一个关键且受调节的处理事件，其中内含子精确从新生的RNA转录物中切除。多达三分之一的遗传疾病突变导致剪接缺陷。这项研究研究了分支点在确定剪接位点选择（3'S）中的作用，在体内使用，分支点对生命周期o内含子的影响。每个前MRNA剪接事件都会产生套索和剪接的外显子连接。虽然关于剪接外显子连接的了解很多，但对套索几乎一无所知。通过绘制人类基因组中的所有分支点，我们正在开放一个全新的分析领域。通过转录本数据识别分支点将促进临床测序数据的解释。除了该数据的内在价值外，该提案的成功完成还将检验一些关于在真核基因加工中体内发生的基本催化和识别的假设。在系统中研究这些中间体将为成千上万的处理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 - 通过剪接的剪接产物创建，并通过脱粒和退化回收。内含子的回收对于补充自由核苷酸的细胞内水平至关重要，并将剪接因子返回到活动剪接中。一些内含子在拼接后作为非编码RNA（NCRNA）具有第二寿命。当我们正在抽样内含子的稳态水平时，我们会深入了解这两个过程。我们的初步数据表明一种新的降解，致力于回收大型内含子。该提议试图通过定义完整的退化途径并探讨某些内含子显得稳定的某些原因，以遵循这一领先。