Investigating Casc3 function in localized mRNA regulation and neuromuscular development

研究 Casc3 在局部 mRNA 调节和神经肌肉发育中的功能

• 批准号: 10448238
• 负责人: Rene Marlene Arvola
• 金额: $ 5.57万
• 依托单位: OHIO STATE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-07-01 至 2023-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10448238
• 关键词: 3' Untranslated Regions; Address; Axon; Biological; Biological Models; Brain; Breast Cancer Patient; Chromosome Mapping; Clustered Regularly Interspaced Short Palindromic Repeats; Complex; Defect; Deposition; Development; Disseminated Malignant Neoplasm; Embryo; Embryonic Development; Eukaryota; Exhibits; Exons; Gene Expression; Genes; Genetic Transcription; Human; Intellectual functioning disability; Introns; Knock-out; Malignant Neoplasms; Mediating; Messenger RNA; Metastatic to; Molecular; Monitor; Morphology; Motor Neurons; Multiprotein Complexes; Mus; Muscle; Nature; Nervous system structure; Neurons; Nonsense-Mediated Decay; Nuclear Export; Organism; Paralysed; Phenotype; Play; Post-Transcriptional Regulation; Predisposition; Proteins; Quality Control; RNA; RNA Decay; RNA Processing; RNA Stability; RNA, Messenger, Splicing; RNA-Binding Proteins; Regulation; Research; Role; Stimulus; Synapses; Testing; Transcript; Zebrafish; cell type; in vivo; insight; lymph nodes; motor neuron development; mutant; nerve stem cell; neurodevelopment; neuromuscular; overexpression; recruit; response; spatiotemporal

Embryonic development requires highly specific spatiotemporal control of gene expression to establish diverse cell types. This control is achieved, in part, by RNA binding proteins that regulate the stability and expression of mRNAs. Due to the dynamic nature of RNA regulation and localization in neurons, neurodevelopment is especially sensitive to perturbations in posttranscriptional regulation. The Exon Junction Complex (EJC) is necessary for brain development through supporting the proliferation of neural progenitors; moreover, EJC core components Rbm8a and Magoh are critical for proper development of motor neurons and muscle during embryogenesis in zebrafish. The EJC is a conserved large multiprotein complex deposited on spliced mRNAs that serves as a regulator of mRNA quality control in higher eukaryotes. The EJC can elicit Nonsense-Mediated Decay (NMD) of improperly spliced mRNAs, but also mRNAs containing 3′ Untranslated Region introns (3UIs). The Cancer Susceptibility Candidate 3 (Casc3) protein is a subunit of the conserved exon-exon junction complex (EJC) that appears to potentiate the decay of a subset of transcripts. Moreover, Casc3 has several documented roles in mRNA localization. While the Casc3 gene was first discovered as a factor that is upregulated in metastatic lymph nodes of breast cancer patients, Casc3 is also highly expressed in the nervous system, and expressed throughout development in zebrafish. This proposal aims to address the biological roles and molecular functions of Casc3 in the control of localized mRNA expression in neuromuscular development. I will utilize zebrafish as a model system to investigate the role of Casc3 in neuromuscular development. I will also characterize Casc3 mechanisms of localization and how they are relevant to specific mRNA features and fates in motor neurons. I hypothesize that Casc3 is necessary neuromuscular development, like other EJC core subunits in vivo, and that Casc3 facilitates localization and decay of 3UI-containing mRNAs in motor neurons. We anticipate that Casc3 function extends to other cellular contexts where it is expressed; for example, dissecting Casc3 functions in a neuronal context may provide insight to how its overexpression could contribute to metastatic cancer. Furthermore, this proposed research will enhance our understanding of the mRNA localization function of Casc3 at the molecular level, which will broadly be relevant to its function in these biological contexts.

胚胎发育需要高度特异性的基因表达时空控制，以建立不同的细胞类型。这种控制部分地通过调节mRNA的稳定性和表达的RNA结合蛋白来实现。由于神经元中RNA调控和定位的动态性质，神经发育对转录后调控的扰动特别敏感。外显子连接复合物（EJC）通过支持神经祖细胞的增殖而对脑发育是必需的;此外，EJC核心组分Rbm 8a和Magoh对于斑马鱼胚胎发生期间运动神经元和肌肉的适当发育至关重要。EJC是一种保守的多蛋白复合物，沉积在剪接的mRNA上，在高等真核生物中作为mRNA质量控制的调节因子。EJC不仅可以引起不正确剪接的mRNA的无义介导的衰变（NMD），还可以引起含有3′非翻译区内含子（3UI）的mRNA的无义介导的衰变。癌症易感性候选者3（Casc 3）蛋白是保守的外显子-外显子连接复合物（EJC）的亚基，其似乎增强转录物子集的衰变。此外，Casc 3在mRNA定位中具有几种记录的作用。虽然Casc 3基因最初被发现是乳腺癌患者转移性淋巴结中上调的一种因子，但Casc 3在神经系统中也高度表达，并在斑马鱼的整个发育过程中表达。该提案旨在解决Casc 3在神经肌肉发育中控制局部mRNA表达的生物学作用和分子功能。我将利用斑马鱼作为模型系统来研究Casc 3在神经肌肉发育中的作用。我还将描述Casc 3的定位机制，以及它们如何与运动神经元中的特定mRNA特征和命运相关。我假设Casc 3是必要的神经肌肉的发展，像其他EJC核心亚基在体内，Casc 3促进本地化和衰减的3UI-在运动神经元中的mRNA。我们预计Casc 3的功能扩展到它表达的其他细胞环境;例如，在神经元环境中解剖Casc 3功能可以提供对其过度表达如何有助于转移性癌症的见解。此外，这项拟议的研究将增强我们在分子水平上对Casc 3的mRNA定位功能的理解，这将与其在这些生物学背景下的功能广泛相关。