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: 10299046
• 负责人: Kexin Xu
• 金额: $ 33.31万
• 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/10299046
• 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; Nuclear Structure; 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 anomaly; 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）的细胞核 - 胞质穿梭是时空的关键决定因素 基因表达的表达。这个过程被多种因素调节，其中之一是添加 甲基部分在氮-6位置（M6A）的腺苷碱基上。新兴证据表明 M6A增强了装饰mRNA的核出口。当前解释M6A辅助mRNA的理论 出口重点是涉及M6A修饰的蛋白质与典型RNA-之间的串扰 将mRNA货物交给经典输出受体的结合衔接蛋白。在这项研究中，我们 已经找到了M6A标记的mRNA的非规范运输轴。 我们的初步数据表明，m6a甲基转移酶复合酶复合物的积分亚基Mettl3， 直接与NUP93相互作用，NUP93是核孔复合物（NPC）的组成部分。 RNA结合能力的能力 METTL3有助于将M6A修饰的mRNA与NUP93连接到核包膜上，这加速了 在主要出口因子NXF1-NXT1的协助下，mRNA货物通过NPC通过。 M6A谱和分级转录组的综合分析已确定了一大批M6A- 带有核出口的mRNA取决于Mettl3-Nup93综合体。有趣的是，这些 mRNA与其他成绩单有区别，因为它们往往具有更长的3'未翻译区域（3'-UTR）， 这与更多的M6A位点和更强的甲基化强度有关。另外，基因本体论 分析揭示了这些转录本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，据报道引起抗类固醇的肾病综合征 （SRN）在人类中，破坏了其与Mettl3的相互作用，这意味着 METTL3-NUP93轴的失调和某些病理条件的发展。被引导 这些初步数据，我们开发了生物系统来监测测试mRNA的M6A状态 并将采用尖端技术，例如单点边缘启用细分显微镜和 表面参考组编辑的工具，以定义Mettl3-Nup93轴在M6A-核输出中的作用 共轭mRNA。我们的具体目标是：（1）。检查mettl3-nup93的相互作用如何促进 M6A标记的mRNA的核出口； （2）。确定哪些成绩单是由 mettl3-nup93轴； （3）。为了阐明为什么这种新的mRNA传输途径在生物学上很重要。 我们提出的研究将影响RNA生物学和表观遗传学领域。从中的发现 研究将有助于描述M6A辅助mRNA导出的新型机制，并提供新的治疗靶标 对于某些人类疾病，例如由NUP93突变引起的肾脏异常。