Nutrient regulation of Alternative splicing and transcription by O-GlcNAcylation

O-GlcNAcylation 对选择性剪接和转录的营养调节

• 批准号: 10535481
• 负责人: Shouling Xu
• 金额: $ 33.92万
• 依托单位: CARNEGIE INSTITUTION OF WASHINGTON, D.C.
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-12-01 至 2024-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10535481
• 关键词: Acinus organ component; Affinity Chromatography; Agriculture; Alternative Splicing; Amino Acids; Animals; Apoptosis; Arabidopsis; Arabidopsis Proteins; Biochemical; Biological Models; Biology; Cell Differentiation process; Chromatin; Complement; Complex; Cues; Data; Development; Dissection; Ensure; Eukaryota; Eukaryotic Cell; Event; Flowers; Fractionation; Gene Expression; Gene Expression Regulation; Genes; Genetic; Genetic Transcription; Genomic approach; Genomics; Germination; Goals; Health; Homologous Gene; Human; Introns; Label; Link; Maps; Mass Spectrum Analysis; Modification; Molecular; Mutagenesis; Names; Nutrient; O-GlcNAc transferase; Pathway interactions; Plant Gene Expression Regulation; Plants; Play; Post-Translational Protein Processing; Productivity; Proteins; Proteomics; RNA; RNA Splicing; RNA-Protein Interaction; Regulation; Regulatory Pathway; Research; Role; Signal Transduction; Site; Specificity; Stress; System; Technology; Testing; Transcriptional Regulation; Transgenic Organisms; Transgenic Plants; biological adaptation to stress; cell growth regulation; crosslink; crosslinking and immunoprecipitation sequencing; experimental study; gene repression; genetic approach; hormonal signals; in vivo; mutant; novel strategies; plant growth/development; posttranscriptional; programs; protein complex; response; sugar; synthetic protein; transcriptome sequencing

The long-term goal of this project is to understand the molecular mechanisms that control gene expression and developmental transitions. While transcription has been extensively studied, the posttranscriptional mechanisms of RNA alternative splicing is much less understood despite of their importance in cellular regulation, human health, and plant growth and development. We have discovered that the Arabidopsis protein AtAcinus is evolutionarily related to but highly divergent from the human Acinus protein, which plays important roles in regulating transcription, RNA alternative splicing, and apoptosis. Our unpublished studies have shown that AtAcinus is modified by O- GlcNAcylation, plays essential role in alternative splicing of a number of genes, many of which encoding key components of signaling and developmental pathways. In particular, our data indicate that AtAcinus play important roles in regulating seed germination and flowering, two major developmental transition in plants. Using a combination of proteomics, genetics, genomic and biochemical approaches in the Arabidopsis model system, we have made tremendous progress in understanding the functions of AtAcinus. Our results support a hypothesis that AtAcinus is controlled by O-GlcNAcylation in response to endogenous and environmental cues, and in turn it regulates key cellular pathways through both transcriptional and posttranscriptional mechanisms. In this proposal, we plan to continue using the combination of proteomic, genomic and genetic approaches to further advance our understanding of Acinus regulatory pathway. We will 1) dissect the molecular functions of AtAcinus, particularly taking advantage of proximity labeling, cross-linking mass spectrometry and biochemical fractionation, CLIP-seq and CLIP-MS technologies to understand how AtAcinus carries out multiple functions (aim 1 and 3); 2) dissect how AtAcinus functions are regulated by post- translational modifications (aim 2). The experiments outlined in this proposal will greatly advance our understanding of the molecular mechanism of RNA alternatively splicing and O-GlcNAcylation and the mechanisms of signal integration at post-transcriptional level. Given the evolutionary conservation of Acinus, this study not only is important for plant biology and agriculture, but also can potentially help us understand fundamental mechanisms of signaling and cellular regulation that are relevant broadly.

该项目的长期目标是了解控制基因的分子机制 表达和发展过渡。虽然转录已经进行了广泛的研究，但 RNA替代剪接的转录后机制尽管有很多知识 在细胞调节，人类健康以及植物生长和发育中的重要性。我们有 发现拟南芥蛋白atacinus在进化上与 人腺泡蛋白在调节转录，RNA替代方面起着重要作用 剪接和凋亡。我们未发表的研究表明，aTacinus是通过O-修改的 Glcnacylation，在许多基因的替代剪接中起着至关重要的作用，其中许多基因 编码信号传导和发育途径的关键组成部分。特别是，我们的数据表明 阿塔奇纳斯在调节种子发芽和开花方面起着重要作用，两个主要 植物的发展过渡。结合蛋白质组学，遗传学，基因组和 拟南芥模型系统中的生化方法，我们在 了解atacinus的功能。我们的结果支持一个假设，即atacinus受到控制 通过O-Glcnacylation响应内源和环境线索，然后调节密钥 通过转录和转录后机制的细胞途径。在此提案中， 我们计划继续使用蛋白质组学，基因组和遗传方法的组合来进一步 促进我们对腺泡调节途径的理解。我们将1）剖析分子功能 atacinus，特别是利用接近标记，交联质谱和 生化分馏，夹式和夹MS技术，以了解Atacinus如何携带 发出多个功能（AIM 1和3）； 2）剖析atacinus函数如何受到后的调节 翻译修改（AIM 2）。该提案中概述的实验将大大提高 我们对RNA的分子机制的理解或O-Glcnacylation替代 以及在转录后级别的信号积分的机制。鉴于进化 抗毒剂的保护，这项研究不仅对植物生物学和农业很重要，而且可以 潜在地帮助我们了解信号传导和细胞调节的基本机制 广泛相关。