Molecular Mechanisms and Signal-Induced Regulation of Alternative Splicing

选择性剪接的分子机制和信号诱导调控

• 批准号: 9913561
• 负责人: KRISTEN W LYNCH
• 金额: $ 58.3万
• 依托单位: UNIVERSITY OF PENNSYLVANIA
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-05-09 至 2021-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9913561
• 关键词: Address; Alternative Splicing; Antigens; Biogenesis; Biological Models; Cell physiology; Cells; Complex; Cues; Disease; Environment; Event; Funding; Gene Expression; Gene Expression Regulation; Genes; Genetic Transcription; Goals; Health; Human; Human body; Immune system; Individual; Lead; Malignant Neoplasms; Metabolism; Methodology; Molecular; Neurons; Pathway interactions; Phosphotransferases; Process; Protein Isoforms; Proteins; RNA; RNA Processing; RNA Splicing; RNA-Binding Proteins; Regulation; Shapes; Signal Pathway; Signal Transduction; Spliceosome Assembly Pathway; System; T-Cell Activation; T-Lymphocyte; cell growth; cell type; detection of nutrient; effector T cell; experimental study; extracellular; immune function; insight; mRNA Precursor; novel; prevent; protein expression; public health relevance; response; tumorigenesis

 DESCRIPTION (provided by applicant): The ultimate goal of this project is to understand the molecular mechanisms by which RNA processing is controlled by signaling pathways, and the impact of this regulation on cellular function. The specific focus of the current funding period is on regulation of alternative pre-mRNA splicing during T cell activation. Alternative splicing is an essential and ubiquitous mechanism of gene regulation, which allows for the diversification and control of protein expression in distinct cell-types or environmental conditions. In particular, itis well established that signal-induced alternative splicing is pervasive during neuronal activity, nutrient sensing, oncogenesis and immune function. However, a mechanistic view of how signaling pathways control alternative splicing remains lacking. T cell activation provides an excellent model system for complex cellular responses, in that multiple signaling pathways are triggered downstream of antigen engagement and act, individually and cooperatively, to induce T cell effector functions. Importantly, several hundred genes are known to undergo alternative splicing in response to T cell activation; however, it remains to be determined how these genes are regulated, which genes are co-regulated by overlapping mechanisms/pathways, and what the ultimate functional consequence is of such regulation. This proposal will address these unanswered questions of signal- induced alternative splicing by leveraging recently developed methodologies and systems to determine: (1) the specific signaling pathways that lead to changes in alternative splicing following antigen stimulation of T cells, and the RNA binding proteins (RBPs) that connect each signaling pathway with their respective splicing targets; (2) the molecular mechanisms by which RBPs and/or cell signaling regulate specific transitions in spliceosome assembly to direct isoform expression; (3) the functional impact of alternative splicing on cell signaling through regulating isoform expression of related kinases; and (4) how regulation of alternative splicing is coordinated with other RNA biogenesis events such as transcription and 3' end processing. Together these studies will provide novel insight regarding the interplay of signaling and splicing in shaping cellular function during T cell activation. Sinc the signaling pathways induced upon T cell activation are also functional in pathways related to cell growth, metabolism and cancer, the insight gained in these studies will be broadly applicable to numerous cellular responses far beyond the immune system. In addition, the studies proposed here will reveal new paradigms regarding the molecular mechanisms by which cells regulate spliceosome assembly, and how RBPs coordinately control splicing along with other steps in RNA processing. Results from these experiments will thus significantly increase understanding of the mechanisms that control alternative splicing, an essential and ubiquitous process for regulating gene expression across all human cell types.

 描述（由申请人提供）：本项目的最终目标是了解RNA加工受信号通路控制的分子机制，以及这种调节对细胞功能的影响。本资金期的具体重点是 调节T细胞活化过程中的可变前体mRNA剪接。选择性剪接是一种 基因调控的基本和普遍存在的机制，允许在不同的细胞类型或环境条件下多样化和控制蛋白质表达。特别是，它是公认的信号诱导的选择性剪接是普遍的神经元活动，营养传感，肿瘤发生和免疫功能。然而，信号通路如何控制可变剪接的机制仍然缺乏。T细胞活化为复杂的细胞应答提供了极好的模型系统，因为多个信号传导途径在抗原接合的下游被触发，并且单独地和协同地起作用以诱导T细胞效应子功能。重要的是，已知数百个基因在响应T细胞活化时经历选择性剪接;然而，仍有待确定这些基因如何被调节，哪些基因通过重叠机制/途径共调节，以及这种调节的最终功能后果是什么。该提案将通过利用最近开发的方法和系统来解决信号诱导的选择性剪接的这些未回答的问题，以确定：（1）在抗原刺激T细胞后导致选择性剪接变化的特异性信号传导途径，以及将每个信号传导途径与其各自的剪接靶点连接的RNA结合蛋白（RBP）;（2）RBP和/或细胞信号传导调节剪接体组装中的特异性转变以指导同种型表达的分子机制：（3）可变剪接通过调节相关激酶的同种型表达对细胞信号传导的功能影响;以及（4）可变剪接的调节如何与其他RNA生物发生事件如转录和3'末端加工协调。这些研究将为T细胞活化过程中信号传导和剪接在塑造细胞功能方面的相互作用提供新的见解。由于T细胞活化后诱导的信号传导途径也在与细胞生长、代谢和癌症相关的途径中起作用，因此这些研究中获得的见解将广泛适用于远远超出免疫系统的许多细胞反应。此外，本文提出的研究将揭示关于细胞调节剪接体组装的分子机制的新范例，以及RBP如何协调控制剪接沿着RNA加工中的其他步骤。因此，这些实验的结果将显着增加对控制选择性剪接的机制的理解，选择性剪接是调节所有人类细胞类型基因表达的一个重要和普遍存在的过程。