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: 10120342
• 负责人: Jeongsik Yong
• 金额: $ 32.82万
• 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/10120342
• 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塑性和替代聚腺苷酸化 项目摘要/摘要 U2AF（U2辅助因子，由U2AF1和U2AF2组成）是必不可少的剪接因子和功能 前MRNA处理期间的3 splice站点选择。众所周知，U2AF形成本构杂化二聚体 对于替代和构成剪接非常重要。但是，初步数据显示MTOR（哺乳动物 雷帕霉素的靶标信号通路控制U2AF2磷酸化的U2AF异构化 - 依赖方式。 U2AF1和U2AF2之间的这些调节的相互作用构成U2AF可塑性。 这种新发现的U2AF可塑性是替代剪接和替代聚腺苷酸化的关键元素。 基于这些发现，可以提出以下中央假设：U2AF可塑性是通往的门户 MTOR调节的转录组重编程。该提议的目标是调查监管 MTOR的U2AF可塑性机理，并了解该U2AF可塑性如何编程转录组 通过关注替代剪接和替代聚腺苷酸化。为此，提出了两个具体目标。 在第一个目标中，将研究MTOR-U2AF可塑性在转录组重编程中的作用。 将进行CRISPR/CAS9介导的基因组工程，以构建细胞模型 以一种或另一种方式使U2AF可塑性偏振。然后将测试这些细胞模型的细胞表型 更改和转录组更改将进行介绍。 U2AF1中的突变在急性髓样中是预后的 白血病和髓样发育不良。这些突变与U2AF可塑性的生理相关性将进行测试，并且 当前的疾病发病机理模型将受到挑战。对于这些任务，新的生物信息学管道将 被开发。在第二个目标中，连接MTOR，U2AF可塑性和组蛋白的调节轴 将解剖生物发生。将确定控制U2AF可塑性的激酶。另外，机制 通过将U2AF可塑性程序替代剪接和替代聚烯基化将被描述。 最后，组蛋白生物发生中MTOR-U2AF塑性介导的替代聚腺苷酸化的结果 将被检查。该项目将共同提高MTOR-对转录组编程的理解 协调的U2AF可塑性，并提出传达细胞外/细胞的机械级联 基因表达程序的环境。它还将挑战当前癌症中U2AF1突变的模型 发病。此外，该项目将通过U2AF建立MTOR与Hisstone生物发生之间的联系 可塑性。