mTOR-regulated U2AF plasticity and alternative polyadenylation

mTOR 调节的 U2AF 可塑性和替代聚腺苷酸化

• 批准号: 10263280
• 负责人: Jeongsik Yong
• 金额: $ 31.62万
• 依托单位: UNIVERSITY OF MINNESOTA
• 依托单位国家: 美国
• 财政年份: 2015
• 资助国家: 美国
• 起止时间: 2015-09-17 至 2024-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10263280
• 关键词: 3' Splice Site; Acute Myelocytic Leukemia; Affect; Alternative Splicing; Biochemical; Biogenesis; Biological; CRISPR/Cas technology; Cell Line; Cell model; Cell physiology; Cells; Chemicals; Complex; Consensus; Coupled; Custom; Data; Deletion Mutation; Disease; Disease model; Dysplasia; Elements; Environment; FRAP1 gene; Gene Expression; Gene Expression Process; Genes; Genome engineering; Goals; Heterodimerization; Histones; Human; Investigation; Length; Libraries; Link; Malignant Neoplasms; Mediating; Messenger RNA; Modeling; Mutation; Myelogenous; Outcome; PC4 Gene; Pathogenesis; Pathway interactions; Phenocopy; Phenotype; Phosphorylation; Phosphotransferases; Physiological; Play; Polyadenylation; Process; Protein Isoforms; Proteome; RNA Interference; RNA Processing; RNA Splicing; Regulation; Renilla Luciferases; Reporter; Role; Signal Pathway; Signal Transduction; Testing; Transcription Coactivator; base; bioinformatics pipeline; bioinformatics tool; cell growth regulation; combinatorial; design; extracellular; interdisciplinary approach; mRNA Precursor; mTOR inhibition; mimetics; mutant; prognostic; programs; transcriptome; transcriptome sequencing; transcriptomics

mTOR-regulated U2AF plasticity and alternative polyadenylation PROJECT SUMMARY/ABSTRACT U2AF (U2 auxiliary factor, comprised of U2AF1 and U2AF2) is an essential splicing factor and functions in 3’splice site selection during pre-mRNA processing. U2AF has been known to form a constitutive heterodimer and is important for alternative and constitutive splicing. However, preliminary data show that mTOR (mammalian target of rapamycin) signaling pathway controls the U2AF heterodimerization by U2AF2 phosphorylation- dependent manner. These regulated interactions between U2AF1 and U2AF2 constitute the U2AF plasticity. This newly discovered U2AF plasticity is a key element in alternative splicing and alternative polyadenylation. Based on these findings, the following central hypothesis can be proposed: the U2AF plasticity is a gateway to mTOR-regulated transcriptome reprogramming. The goals of this proposal are to investigate the regulatory mechanism of U2AF plasticity by mTOR and understand how this U2AF plasticity programs the transcriptome by focusing on alternative splicing and alternative polyadenylation. To this end, two specific aims are proposed. In the first aim, the role of mTOR-U2AF plasticity in transcriptome reprogramming will be investigated. CRISPR/Cas9-mediated genome engineering will be conducted to build up cell models which will constitutively polarize the U2AF plasticity in one way or the other. These cell models will be then tested for cell phenotypic changes and the transcriptomic changes will be profiled. Mutations in U2AF1 are prognostic in acute myeloid leukemia and myeloid dysplasia. Physiological relevance of these mutations to U2AF plasticity will be tested and a current model for disease pathogenesis will be challenged. For these tasks, a new bioinformatic pipeline will be developed. In the second aim, the regulatory axis that connects mTOR, U2AF plasticity, and histone biogenesis will be dissected. A kinase(s) that controls the U2AF plasticity will be identified. Also, the mechanism by which the U2AF plasticity programs alternative splicing and alternative polyadenylation will be delineated. Finally, the outcome of mTOR-U2AF plasticity-mediated alternative polyadenylation in the histone biogenesis will be examined. Together, this project will advance the understanding of transcriptome programming by mTOR- coordinated U2AF plasticity and suggest mechanistic cascades that communicate extracellular/cellular environments to gene expression programs. It will also challenge a current model of U2AF1 mutations in cancer pathogenesis. Moreover, this project will establish a link between mTOR and histone biogenesis through U2AF plasticity.

MTOR调节的U2AF可塑性和交替多聚腺苷 项目摘要/摘要 U2AF(U2辅助因子，由U2AF1和U2AF2组成)是一种重要的剪接因子，在 3‘剪接位点在前mRNA加工过程中的选择。已知U2AF形成构造性杂二聚体 并且对于可选的和构成的剪接是重要的。然而，初步数据显示，mTOR(哺乳动物 雷帕霉素靶标)信号通路通过U2AF2磷酸化调控U2AF异源二聚。 依赖的态度。U2AF1和U2AF2之间的这些调控相互作用构成了U2AF的可塑性。 这种新发现的U2AF可塑性是选择性剪接和选择性多聚腺苷酸化的关键因素。 基于这些发现，可以提出以下中心假设：U2AF的可塑性是 MTOR调控的转录组重新编程。这项提案的目标是调查监管机构 通过mTOR研究U2AF可塑性的机制并了解这种U2AF可塑性如何对转录组进行编程 通过专注于选择性剪接和选择性聚腺苷酸化。为此，提出了两个具体目标。 在第一个目标中，将研究mTOR-U2AF可塑性在转录组重新编程中的作用。 将进行CRISPR/Cas9介导的基因组工程来建立细胞模型，该模型将构成 以这样或那样的方式极化U2AF的可塑性。然后将对这些细胞模型进行细胞表型测试 改变和转录改变将被描述。U2AF1基因突变可预测急性髓系白血病的预后 白血病和髓系异常增殖症。这些突变与U2AF可塑性的生理相关性将被测试和 目前的疾病发病机制模型将受到挑战。对于这些任务，一条新的生物信息学管道将 被开发出来。在第二个目标中，连接mTOR、U2AF可塑性和组蛋白的调节轴 生物发生学将被解剖。控制U2AF可塑性的激酶(S)将被确定。此外，该机制 其中，U2AF可塑性程序，选择性剪接和选择性多聚腺苷将被描绘。 最后，mTOR-U2AF可塑性介导的交替多聚腺苷化在组蛋白生物发生中的结果 将会被检查。总之，该项目将促进对mTOR转录组编程的理解- 协调的U2AF可塑性，并提出了沟通细胞外/细胞的机械级联 环境对基因表达程序的影响。它还将挑战目前癌症中U2AF1突变的模型 发病机制。此外，该项目将通过U2AF在mTOR和组蛋白生物发生之间建立联系 可塑性。