Protein RNA Rearrangements in the Spliceosome

剪接体中蛋白质 RNA 重排

基本信息

批准号：
10367405
负责人：
CHARLES C QUERY
金额：
$ 55.44万
依托单位：
ALBERT EINSTEIN COLLEGE OF MEDICINE
依托单位国家：
美国
项目类别：
财政年份：
1999
资助国家：
美国
起止时间：
1999-08-01 至 2026-01-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10367405
关键词：
3&apos Splice Site ATP phosphohydrolase Alleles Binding Binding Sites Biochemistry Bioinformatics Biological Biology Catalysis Cell Nucleus Cells Chemicals Clustered Regularly Interspaced Short Palindromic Repeats Code Complex Cryoelectron Microscopy Data Data Set Defect Development Disease Dysmyelopoietic Syndromes Enzymes Event Excision Exons Gambling Gene Expression Genes Genetic Genetic Screening Genetic Transcription Genome Glioblastoma Goals Human Human Cell Line Individual Inositol Introns Investigation Joining Exons Kinetics Ligands Link Malignant Neoplasms Malignant neoplasm of urinary bladder Mammalian Cell Mediating Methods Modification Molecular Conformation Mutation Nucleotides Organoids Pathway interactions Phenotype Phytic Acid Polyadenylation Polyphosphates Process Property Protein Isoforms Proteins RNA RNA Splicing Reaction Recurrence Reverse engineering Role SAGA Site Small Nuclear RNA Small Nuclear Ribonucleoproteins Spinal Muscular Atrophy Spliceosome Assembly Pathway Spliceosomes Structure System Testing The Cancer Genome Atlas Transcription Elongation Trinucleotide Repeats Tweens U1 small nuclear RNA U2 Small Nuclear Ribonucleoprotein U2 small nuclear RNA U5 small nuclear RNA Variant Yeasts base bioinformatics pipeline experimental study genetic variant genome-wide human disease improved in silico interest mRNA Precursor mammalian genome mutant public health relevance small molecule stem transcriptome sequencing whole genome yeast genetics

项目摘要

ABSTRACT Excision of introns from precursor messenger RNA by the spliceosome is a critical step in almost all human gene expression. This process is highly regulated, integrally linked with the transcription of genes and other processing events, such as polyadenylation and nucleotide modification. The mechanism by which the spliceosome recognizes the exact sites for the chemical events and how the reactions are catalyzed are not well understood. The long-term goals of this project are to understand interactions and rearrangements between spliceosome components and the RNA ligands that are substrates for the catalytic reactions. Ample evidence argues for multiple rearrangements of factors and multiple recognition events at the branch site. Investigation of these events — which are not understood mechanistically — will elucidate interactions and rearrangements among core components and may serve as a paradigm for rearrangements in the spliceosome and in other RNP machines. This proposal focuses on mechanisms by which spliceosomal dynamics impact splicing fidelity. The Aims of this project focus on two newly-discovered aspects of the spliceosome and snRNAs: (1) We will investigate the role of inositol hexakisphosphate (IP6) in spliceosome function. This small molecule was recently identified unexpectedly by cryo-EM as part of splicing complexes. No biochemistry or genetics has so far explored its importance. We use mutants in the IP6 biosynthetic pathway to explore its contribution to splicing and whether inositol polyphosphate (IPP) isoform levels may influence splice events. (2) One hole in our current understanding of RNA biology is the role of snRNA mutations in disease. Two mutations were recently described in the 5'-end of U1 snRNA, associated with glioblastoma (GBM) and other cancers. We will use our in silico snRNA genome to interrogate TCGA whole genome datasets (WGS) and investigate, in particular, U2 snRNA mutations that we find highly enriched in bladder cancer and bladder cancer organoid lines.

抽象的剪接体从前体信使 RNA 中切除内含子是几乎所有研究中的关键步骤。人类基因表达受到高度调控，与基因的转录密切相关。其他加工事件，例如聚腺苷酸化和核苷酸修饰。剪接体识别化学事件的确切位点的机制以及如何该项目的长期目标是了解所催化的反应。剪接体成分和底物 RNA 配体之间的相互作用和重排充足的证据支持因素的多重重排和多重反应。对分支机构现场的识别事件进行调查——这些事件尚不清楚。机械地——将阐明核心组件之间的相互作用和重新排列，并可以作为该提案重点关注剪接体和其他 RNP 机器中的重排范例。剪接体动力学影响剪接保真度的机制。该项目的目标集中于剪接体和 snRNA 的两个新发现的方面： (1) 我们将研究肌醇六磷酸（IP6）这种小分子在剪接体功能中的作用。最近被冷冻电镜意外地鉴定为剪接复合物的一部分，无生物化学或遗传学。到目前为止，我们已经利用 IP6 生物合成途径中的突变体来探索其贡献。剪接以及肌醇多磷酸 (IPP) 同工型水平是否可能影响剪接事件 (2) 一孔。我们目前对 RNA 生物学的理解是 snRNA 突变在疾病中的作用。最近描述了 U1 snRNA 的 5' 端与胶质母细胞瘤 (GBM) 和其他癌症相关。使用我们的计算机 snRNA 基因组来询问 TCGA 全基因组数据集 (WGS) 并进行研究特别是，我们发现膀胱癌和膀胱癌类器官中高度富集的 U2 snRNA 突变线。