METTL3-NUP93 interaction facilitates the nuclear export of m6A-modified mRNAs

METTL3-NUP93 相互作用促进 m6A 修饰 mRNA 的核输出

• 批准号: 10461179
• 负责人: Kexin Xu
• 金额: $ 33.33万
• 依托单位: UNIVERSITY OF TEXAS HLTH SCIENCE CENTER
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-08-10 至 2026-07-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10461179
• 关键词: 3' Untranslated Regions; Adaptor Signaling Protein; Adenosine; Affect; Apoptosis; Articulation; Binding; Biological; Biological Assay; Biology; CRISPR/Cas technology; Cell Cycle Progression; Cell Nucleus; Cell Proliferation; Cell physiology; Cells; Chronic Kidney Failure; Complex; Cytoplasm; DNA Sequence Alteration; Data; Defect; Development; Enzymes; Epigenetic Process; Fibrinogen; Fractionation; Gene Expression; Genes; Hand; Health; Human; Immunoprecipitation; Kidney; Kinetics; Knowledge; Label; Ligation; Link; Mediating; Messenger RNA; Methylation; Methyltransferase; Microscopy; Modification; Molecular; Monitor; Movement; Mutation; Nitrogen; Nuclear; Nuclear Envelope; Nuclear Export; Nuclear Pore Complex; Nuclear Pore Complex Proteins; Ontology; Pathogenesis; Pathologic; Pathway interactions; Patients; Pattern; Polyribosomes; Positioning Attribute; Process; Production; Proteins; Proteomics; RNA; RNA Binding; RNA Transport; Regulation; Reporting; Research; Resolution; Role; Sampling; Signal Transduction; Site; Speed; Steroid-resistant idiopathic nephrotic syndrome; System; Technology; Testing; Transcript; Untranslated Regions; base; biological systems; clinically relevant; clinically significant; congenital anomalies of the kidney; density; epitranscriptome; epitranscriptomics; human disease; insight; knock-down; mutant; new therapeutic target; novel; receptor; recruit; scaffold; spatiotemporal; stem; theories; tool; transcriptome; transcriptome sequencing

PROJECT ABSTRACT The nucleus-cytoplasm shuttling of messenger RNAs (mRNAs) is a key determinant in the spatiotemporal articulation of gene expression. This process is finely tuned by multiple factors, one of which is the addition of a methyl moiety onto the adenosine base at the nitrogen-6 position (m6A). Emerging evidence suggests that m6A enhances the nuclear export of decorated mRNAs. Current theories explaining m6A-assisted mRNA export focus on the crosstalk between the protein enzymes involved in m6A modification and the typical RNA- binding adaptor proteins that hand the mRNA cargoes over to the classical export receptors. In this study, we have found a noncanonical transport axis for m6A-marked mRNAs. Our preliminary data have shown that METTL3, an integral subunit of the m6A methyltransferase complex, directly interacts with NUP93, a component of the nuclear pore complex (NPC). RNA-binding capacity of METTL3 helps link the m6A-modified mRNAs to NUP93 on the nuclear envelope, which accelerates the passage of mRNA cargoes through the NPCs with the assistance of the principal export factor NXF1-NXT1. Integrative analysis of the m6A profile and fractionation transcriptome has identified a large group of m6A- harboring mRNAs whose nuclear export is dependent on the METTL3-NUP93 complex. Interestingly, these mRNAs are distinguished from other transcripts, as they tend to have longer 3' untranslated regions (3'-UTR), which is associated with more m6A sites and stronger methylation intensities. In addition, Gene Ontology analysis has revealed important functions of these transcripts in cellular physiology. In this regard, we found that the genetic mutations on NUP93, which have been reported to cause steroid-resistant nephrotic syndrome (SRNS) in humans, disrupts its interaction with METTL3, implying a potential association between dysregulation of METTL3-NUP93 axis and development of certain pathological conditions. Being guided by these preliminary data, we have developed biological systems to monitor the m6A status of test mRNA species and will employ cutting-edge technologies, such as single-point edge-excitation subdiffraction microscopy and epitranscriptome-editing tools, to define the role of METTL3-NUP93 axis in the nuclear export of m6A- conjugated mRNAs. Our Specific Aims are: (1). To examine how the METTL3-NUP93 interaction facilitates the nuclear export of m6A-marked mRNAs; (2). To determine which transcripts are specifically regulated by the METTL3-NUP93 axis; (3). To elucidate why this new mRNA transport pathway is biologically important. Our proposed research will impact upon the fields of RNA biology and epigenetics. Findings from this study will help delineate a novel mechanism of m6A-assisted mRNA export and offer new therapeutic targets for certain human diseases, such as kidney abnormalities that are caused by NUP93 mutations.

项目摘要 信使RNA（mRNA）的核质穿梭是时空变化的关键决定因素。 基因表达的表达。该过程通过多种因素进行微调，其中之一是添加 甲基部分在氮-6位（m6A）的腺苷碱基上。新出现的证据表明 m6A增强修饰的mRNA的核输出。当前解释m6A辅助mRNA的理论 出口集中在m6A修饰中涉及的蛋白酶和典型的RNA之间的串扰， 结合衔接蛋白，将mRNA货物传递给经典的输出受体。本研究 已经发现了m6A标记mRNA的非经典转运轴。 我们的初步数据表明，胃L3，m6A甲基转移酶复合物的一个完整亚基， 直接与NUP93相互作用，NUP93是核孔复合物（NPC）的组分。RNA结合能力 胃L3帮助将m6A修饰的mRNA连接到核膜上的NUP93，这加速了细胞的增殖。 mRNA货物在主要输出因子NXF1-NXT 1的帮助下通过NPC。 m6A谱和分级转录组的综合分析已经鉴定了一大组m6A- 含有依赖于胃L3-NUP93复合物的核输出的mRNA。有趣的是，这些 mRNA与其他转录物不同，因为它们倾向于具有较长的3 '非翻译区（3'-UTR）， 这与更多的m6A位点和更强的甲基化强度有关。此外，Gene Ontology 分析揭示了这些转录物在细胞生理学中的重要功能。在这方面，我们发现 NUP93上的基因突变，据报道会导致类固醇耐药性肾病综合征， （SRNS），破坏其与胃L3的相互作用，这意味着 胃L3-NUP93轴的失调和某些病理状况的发展。的引导 根据这些初步数据，我们开发了生物系统来监测测试mRNA种类的m6A状态 并将采用尖端技术，如单点边缘激发亚衍射显微镜， epitranscriptome编辑工具，以确定胃L3-NUP93轴在m6A核输出中的作用- 结合的mRNA。我们的具体目标是：（1）。为了研究胃L3-NUP93相互作用如何促进胃蛋白酶活性， m6A标记的mRNA的核输出;（2）.为了确定哪些转录本是由 胃L3-NUP93轴;（3）.为了阐明为什么这种新的mRNA转运途径在生物学上是重要的。 我们提出的研究将影响RNA生物学和表观遗传学领域。时发现的问题 这项研究将有助于阐明m6A辅助mRNA输出的新机制，并提供新的治疗靶点 对于某些人类疾病，如NUP93突变引起的肾脏异常。