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剪接是一个关键且受调节的处理事件，其中内含子是 精确地从新生的RNA转录本中切除。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 确定剪接站点选择（3's）在体内使用，效果分支点也具有 关于内含子的生命周期。每个前MRNA剪接事件都会产生套索和剪接外显子 交界处。虽然关于剪接外显子连接的知之甚少，但几乎一无所知 关于套索。通过映射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。通过成绩单数据识别分支点将 促进临床测序数据的解释。除了固有值 这些数据，该提案的成功完成将检验有关该提案的一些假设 在真核基因的处理中，体内发生的基本催化和识别。 在系统中研究这些中间体将使生化水平的理解给 成千上万的处理事件。此外，每个内含子套房都有生命周期 - 通过剪接转录的产品创建，通过脱节和退化回收。这 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 将剪接因子归还为主动剪接体。一些内含子在剪接后具有第二次生命 作为非编码RNA（NCRNA）。由于我们正在抽样内含子的稳态水平，因此我们获得了洞察力 进入这两个过程。该提议试图通过探索一些原因来遵循这一领导 为什么某些内含子显得稳定。