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（由适用提供）：该项目的最终目标是了解通过信号通路控制RNA处理的分子机制，以及该调节对细胞功能的影响。当前资助期的具体重点是 在T细胞活化过程中调节替代前MRNA剪接。替代剪接是 基因调节的必需和无处不在的机制，可以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，在神经元活性，营养敏感性，肿瘤发生和免疫学功能期间，信号诱导的替代剪接具有普遍性。但是，仍然缺乏信号通路如何控制替代剪接的机械视图。 T细胞激活为复杂的细胞响应提供了出色的模型系统，因为多个信号通路是抗原参与度的下游触发的，并且在单独和合作上触发了诱导T细胞效应子功能。重要的是，已知几百个基因会响应T细胞激活而经历替代剪接。但是，尚待确定如何调节这些基因，哪些基因是由重叠的机制/途径共同调节的，以及最终功能后果是这样调节的。该提案将通过利用最近开发的方法和系统来解决这些未解决的问题，以确定：（1）特定的信号传导途径，导致T细胞抗原刺激T细胞以及RNA结合蛋白（RBP）的替代剪接发生变化，以及将每个信号通路与其相应的剪接靶标连接起来； （2）RBP和/或细胞信号传导调节剪接体组装中的特定过渡以导向同工型表达的分子机制； （3）替代剪接对通过相关激酶的细胞信号传导的功能影响； （4）如何与其他RNA生物发生事件（例如转录和3'末端加工）协调替代剪接的调节。这些研究将共同​​提供有关信号传导和剪接在T细胞激活过程中塑造细胞功能中的相互作用的新见解。 SINC在T细胞激活时诱导的信号传导途径在与细胞生长，代谢和癌症有关的途径中也起作用，这些研究中获得的洞察力将广泛适用于远远超出免疫系统的众多细胞反应。此外，此处提出的研究将揭示有关细胞调节剪接体组装的分子机制的新范式，以及RBP如何协调控制剪接以及RNA处理中的其他步骤。因此，这些实验的结果将显着增加对控制替代剪接的机制的理解，这是确定所有人类细胞类型基因表达的必不可少的和无处不在的过程。