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: 10251555
• 负责人: William G Fairbrother
• 金额: $ 2.53万
• 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 2023-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10251555
• 关键词: Antibodies; Area Analyses; Award; Binding; Biochemical; Biochemistry; Biological Assay; Brain Stem; Catalysis; Cells; Cellular Assay; Chemicals; Chromatin; DNA; DNA Polymerase II; Data; Defect; Disease; Enzymes; Event; Exercise; Exons; Failure; Genes; Genetic; Genetic Transcription; Genome; Genomic approach; Heritability; Human; Human Genome; Immunity; In Vitro; Introns; Lead; Life; Life Cycle Stages; Maps; Messenger RNA; Methods; Modification; Molecular; Mutation; Names; Natural Immunity; Nucleotides; Paper; Parents; Pathway interactions; Process; RNA; RNA Splicing; RNA Stability; RNA-Binding Proteins; Reagent; Recycling; Research; Role; Sampling; Site; Small Nuclear Ribonucleoproteins; Spliceosomes; Structural Biochemistry; Structure; System; Testing; Tissues; Transcript; Untranslated RNA; Viral; Virus; Work; clinical sequencing; deep sequencing; experience; improved; in vivo; insight; lariat debranching enzyme; mRNA Precursor; mutant; novel; preservation; research study; scale up; sensor; small molecule inhibitor; structural biology

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 of 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 of clinical sequencing data. In addition to the intrinsic value of this data, the successful completion 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 active 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. This proposal seeks to follow this lead by exploring some of the reasons why certain introns appear stabilized.

前 mRNA 剪接是一个关键的、受调控的加工事件，内含子在其中 从新生RNA转录本中精确切除。多达三分之一的遗传性疾病 突变导致剪接缺陷。本研究研究分支点在 确定剪接位点选择（3'sss）在体内的应用以及分支点的影响 内含子的生命周期。每个前 mRNA 剪接事件都会创建套索和剪接外显子 交界处。虽然人们对剪接外显子连接了解很多，但几乎一无所知 关于套索。通过绘制人类基因组中的所有分支点，我们正在开辟一个 全新的分析领域。通过转录数据识别分支点将 促进临床测序数据的解释。除了内在价值之外 根据这些数据，该提案的成功完成将检验一些关于 真核基因加工过程中体内发生的基本催化和识别。 在系统范围内研究这些中间体将带来生化水平的理解 数十万个处理事件。此外，每个内含子套索都有一个生命周期—— 通过拼接转录产物而产生，通过脱支和降解而回收。这 内含子的回收对于补充细胞内游离核苷酸的水平至关重要 将剪接因子返回到活性剪接体中。一些内含子在剪接后有第二次生命 作为非编码 RNA (ncRNA)。当我们对内含子的稳态水平进行采样时，我们获得了洞察力 进入这两个过程。该提案旨在通过探索一些原因来追随这一趋势 为什么某些内含子看起来稳定。