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

基本信息

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
项目类别：
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

项目摘要

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.

项目概要复杂生物体的发育需要细胞类型和组织的多样化。干细胞等细胞细胞和胚胎必须极化以分离细胞命运决定因素，然后不对称分裂产生功能不同的细胞。未能建立细胞极性是许多癌症和癌症的一个标志发育缺陷。因此，了解细胞极性网络如何耦合具有重要意义通过细胞周期，深入了解特定细胞类型在发育过程中如何形成。 PAR 蛋白是在许多生物体中控制细胞极性正确建立的特殊蛋白。为了例如，小鼠胚胎干细胞通过将 PAR 蛋白定位于顶端或底端来建立顶端-基底极性。基底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，通过形成相互排斥的 PAR 皮质域来建立前后极性。然而 PAR 蛋白是如何调节的尚不完全清楚。最近的研究表明，秀丽隐杆线虫需要细胞周期激酶 Aurora A 来启动极化合子。这些研究表明 Aurora A 表现出动态定位并在不同阶段发挥作用细胞周期来及时、精确地控制皮质 PAR 蛋白的募集。然而， Aurora A 调节 PAR 蛋白不对称性的确切机制仍不清楚。该提案的总体目标是剖析不同的 Aurora A 池如何传输信号促进及时、精确的细胞极化，并确定 Aurora A 在极化过程中的作用是否在其他极化细胞中保守。中心假设是Aurora A在不同的时间和地点起作用调节极性蛋白组织，从而协调极性与细胞周期。这个假设将使用独特的生化方法进行测试，以量化单个细胞、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。目标实验 1 将鉴定线虫中的 Aurora A 底物并测试 Aurora A 活性的丧失如何影响 PAR 蛋白本地化和复杂的组装。目标 2 将确定需要 Aurora A 活动的地点和时间协调 PAR 蛋白，利用光遗传学操纵 Aurora A 空间调控并进行商业化可用Aurora A抑制剂来控制Aurora A活性。目标 3 将评估 Aurora A 活性是否小鼠胚胎干细胞顶端-基底极性建立过程中所需的。总的来说，这些数据将揭示 Aurora A 如何协调极性蛋白动力学和复杂组装，并提供极性如何建立并与细胞周期同步的基础知识。