How a cell cycle kinase, Aurora A, regulates polarity proteins organization

细胞周期激酶 Aurora A 如何调节极性蛋白组织

• 批准号: 10389697
• 负责人: Nadia Ingabire Manzi
• 金额: $ 3.75万
• 依托单位: UNIVERSITY OF TEXAS AT AUSTIN
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-06-01 至 2025-05-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10389697
• 关键词: Address; Affect; Apical; Arthropods; Binding; Biochemical; Caenorhabditis elegans; Cell Cycle; Cell Cycle Stage; Cell Polarity; Cell Separation; Cell division; Cells; Centrosome; Complex; Congenital Abnormality; Coupled; Data; Defect; Development; Embryo; Ensure; Event; Exhibits; Failure; Human; Image; Knowledge; Light; Location; Malignant Neoplasms; Measures; Meiosis; Microscopy; Mitosis; Modeling; Molecular; Mus; Mutation; Nematoda; Organism; Phenotype; Phosphorylation; Phosphorylation Site; Phosphotransferases; Photoreceptors; Positioning Attribute; Protein Dynamics; Proteins; RNA Interference; Regulation; Resolution; Role; Signal Transduction; Site; System; Testing; Therapeutic; Time; Tissues; Work; cancer type; cell type; daughter cell; dimer; embryonic stem cell; experimental study; genome editing; inhibitor; insight; mimetics; mutant; optogenetics; polarized cell; protein complex; protein protein interaction; public health relevance; recruit; segregation; single molecule; stem cells; zygote

Project Summary The development of complex organisms requires diversification of cell types and tissues. Cells such as stem cells and embryos must polarize in order to segregate cell fate determinants, and then divide asymmetrically to generate functionally distinct cells. Failure to establish cell polarity is a hallmark of many cancers and developmental defects. Therefore, it is of great importance to understand how cell polarity network is coupled with the cell cycle, to get insights into how specific cell types are formed during development. PAR proteins are specialized proteins that govern proper establishment of cell polarity in many organisms. For example, mouse embryonic stem cells establish apical-basal polarity by localizing PAR proteins to the apical or basal domains to allow the segregation of cell fate determinants. Another example is the Caenorhabditis elegans zygote, which establishes antero-posterior polarity by forming mutually exclusive PAR cortical domains. However how PAR proteins are regulated is not fully understood. Recent studies have revealed that a cell cycle kinase, Aurora A, is required to initiate polarization in a C. elegans zygote. These studies showed evidence that Aurora A exhibits dynamic localization and acts at different stages of the cell cycle to control cortical recruitment of PAR proteins in a timely and precise manner. However, the exact mechanisms by which Aurora A modulates PAR protein asymmetries remain obscure. The overreaching objectives of this proposal are to dissect how different pools of Aurora A transmit signals that promote timely and precise cell polarization, and to determine if the role of Aurora A during polarization is conserved in other polarizing cells. The central hypothesis is that Aurora A works at different times and places to modulate polarity proteins organizations, thereby coordinating polarity with the cell cycle. This hypothesis will be tested using a unique biochemical approach for quantifying protein-protein interactions at a single cell, single molecule level, in combination with high-resolution microscopy and targeted genome editing. Experiments in Aim 1 will identify Aurora A substrates in C. elegans and test how loss of Aurora A activity impacts PAR proteins localization and complex assembly. Aim 2 will determine where and when Aurora A activity is required to coordinate PAR proteins, using optogenetics to manipulate Aurora A spatial regulation and commercially available Aurora A inhibitors to control the Aurora A activity. Aim 3 will assess whether Aurora A activity is required during apical-basal polarity establishment of mouse embryonic stem cells. Collectively, these data will reveal how Aurora A is positioned to orchestrate polarity proteins dynamics and complex assembly and provide fundamental knowledge of how polarity is established and synchronized with the cell cycle.

项目摘要 复杂的生物的发展需要细胞类型和组织的多样化。细胞，例如茎 细胞和胚胎必须极化才能隔离细胞命运决定因素，然后不对称地分裂 生成功能上不同的细胞。无法建立细胞极性是许多癌症的标志 发展缺陷。因此，了解如何耦合细胞极性网络非常重要 通过细胞周期，以了解开发过程中特定细胞类型的形成方式。 PAR蛋白是专门的蛋白质，可在许多生物体中正确建立细胞极性。为了 例如，小鼠胚胎干细胞通过将PAR蛋白定位到顶端或 基础结构域允许细胞命运决定因素的分离。另一个例子是秀丽隐杆线虫 合子通过形成互斥的皮质域来建立前后极性。然而 如何调节PAR蛋白是未完全理解的。 最近的研究表明，在秀丽隐杆线虫中启动极化需要细胞周期激酶Aurora A 合子。这些研究表明，Aurora A表现出动态定位并在不同阶段起作用的证据 细胞周期以及时，精确的方式控制PAR蛋白的皮质募集。但是， Aurora A调节PAR蛋白不对称的确切机制仍然晦涩。 该提案的过度目标是剖析不同的极光池如何传递信号，表明 促进及时，精确的细胞极化，并确定极光化在极化过程中的作用是否为 在其他偏振细胞中保守。中心假设是Aurora A在不同时间和地方工作 调节极性蛋白质组织，从而将极性与细胞周期进行协调。这个假设将会 使用独特的生化方法进行测试，以量化单个细胞的蛋白质 - 蛋白质相互作用 分子水平，结合高分辨率显微镜和靶向基因组编辑。 AIM中的实验 1将识别秀丽隐杆线虫中的极光A底物，并测试Aurora A活动的损失如何影响PAR蛋白 本地化和复杂组装。 AIM 2将确定需要Aurora的何时何时需要活动 协调PAR蛋白，使用光遗传学来操纵Aurora A空间调节和商业 可用的Aurora A抑制剂来控制Aurora A活性。 AIM 3将评估Aurora A活动是否是 在顶端极性建立小鼠胚胎干细胞期间需要的。总的来说，这些数据将 揭示Aurora a的定位是如何协调极性蛋白动力学和复杂组装并提供 关于如何建立极性并与细胞周期同步的基本知识。