Investigating the molecular mechanisms of early neuronal morphogenesis

研究早期神经元形态发生的分子机制

• 批准号: 10545077
• 负责人: Jernej Murn
• 金额: $ 32.66万
• 依托单位: UNIVERSITY OF CALIFORNIA RIVERSIDE
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-01-01 至 2027-01-01
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10545077
• 关键词: Affect; Binding; Binding Sites; Biochemical; Biological Assay; Bipolar Neuron; Cell Shape; Cell physiology; Cells; Cellular Morphology; Code; Coupled; Cues; Development; Disease; Dissociation; Endowment; Event; FRAP1 gene; Foundations; Gene Expression; Gene Structure; Goals; Homeostasis; Individual; Instruction; Length; Life; Link; Locales; Mammalian Cell; Mediating; Messenger RNA; Microscopy; Modality; Molecular; Morphogenesis; Morphology; Mutagenesis; Nature; Neuroglia; Neurons; Organism; Phosphorylation; Phosphotransferases; Play; Poly(A) Tail; Post-Transcriptional Regulation; Post-Translational Protein Processing; Process; Proteins; Proteomics; RNA; RNA Binding; RNA Recognition Motif; RNA-Binding Proteins; Reporter; Role; Series; Shapes; Site; Somatic Cell; Structure; Supporting Cell; Transcript; Translations; cell type; comparative; design; experimental study; human disease; in vivo; insight; mRNA Translation; nervous system disorder; polarized cell; programs; protein complex; protein protein interaction; recruit; ribosome profiling; spatiotemporal

SUMMARY Cellular morphology is one of the most distinctive features of somatic cells in multicellular organisms and is intimately linked with cellular function. How neurons and other polarized cells commit to their morphologies is poorly understood, but spontaneous morphogenesis of dissociated cells in culture suggest that the basic instructions for morphology are often intrinsically encoded. We previously identified a sequence-specific RNA- binding protein, Unkempt, as a factor that is essential for the establishment of the early neuronal morphology and as a protein that is capable of endowing a similar shape to cells of nonneuronal origin. Unkempt recognizes a unique binding motif predominantly within coding regions mRNAs the translation of which it suppresses. It is unclear how Unkempt regulates translation, and whether translation or another functional modality of Unkempt is critical to its induction of cell polarization. Here, we seek to solve this problem by deciphering the mechanistic basis of Unkempt-driven remodeling of cellular shape. Our preliminary studies indicate that in the broad protein-protein interaction network of Unkempt, the interaction between Unkempt’s low-complexity domain (LCD) and the CCR4-NOT complex is exquisitely required for the induction of cell morphogenesis. We propose three specific aims to investigate the molecular and cellular roles of the CCR4- NOT complex as a critical effector of Unkempt-controlled cell morphogenesis. First, we will investigate the nature of Unkempt – CCR4-NOT interactions and their impact on the fate of the targeted messages, focusing in particular on their poly(A) tail length, stability, and translation. Second, we will interrogate the recruitment and function of CCR4-NOT in Unkempt-induced cell morphogenesis. Third, we will determine the effect of post-translational modifications of Unkempt on its interactions with the CCR4-NOT complex and RNA, as well as its control of local protein translation. This study will shed light on the molecular underpinnings of the early neuronal morphogenesis and contribute to our general understanding of the cues that control cellular morphology in development and disease.

概括 细胞形态是多细胞生物中体细胞最独特的特征之一，是 与细胞功能密切相关。神经元和其他极化细胞如何致力于其形态 知之甚少，但是培养中解离细胞的赞成形态发生表明基本 形态学的说明通常是内在编码的。我们先前鉴定了一个序列特异性的RNA- 结合蛋白质，蓬松，是建立早期神经元形态至关重要的因素 作为一种能够赋予与非神经元来源细胞相似形状的蛋白质。蓬头垢面 在编码区域中，主要识别一个独特的绑定基序mRNA中的翻译 抑制。目前尚不清楚如何调节翻译，以及翻译还是其他功能 露天的模态对于其诱导细胞极化至关重要。在这里，我们试图通过 解读细胞形状的不受欢迎驱动重塑的机械基础。我们的初步研究 表明在宽阔的蛋白质 - 蛋白质相互作用网络中 低复杂结构域（LCD）和CCR4-不合适的复合物是诱导细胞的确切所必需的 形态发生。我们提出了三个特定的目的，以研究CCR4-的分子和细胞作用 作为不受欢迎的控制细胞形态发生的关键效应因子并不复杂。首先，我们将调查 蓬松的性质 - CCR4不相互作用及其对目标信息命运的影响，重点 尤其是其poly（a）尾巴长度，稳定性和翻译。其次，我们将询问招聘 CCR4的功能 - 不在不受欢迎的细胞形态发生中。第三，我们将确定 在与CCR4的相互作用上，透明的翻译修饰也是不合理的，也不是RNA 作为对局部蛋白质翻译的控制。这项研究将阐明早期的分子基础 神经元形态发生，并有助于我们对控制细胞的线索的一般理解 发育和疾病的形态。