Dissecting the role of Aurora A kinase in patterning the cell cortex during cytokinesis.

剖析 Aurora A 激酶在胞质分裂过程中细胞皮质模式化中的作用。

• 批准号: 10634513
• 负责人: Aleesa Schlientz
• 金额: $ 6.95万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN DIEGO
• 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 2024-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10634513
• 关键词: Actins; Actomyosin; Address; Anaphase; Animals; Biological Assay; Biological Models; Caenorhabditis elegans; Cell division; Cell membrane; Cells; Centrosome; Chromosome Segregation; Chromosomes; Cleaved cell; Clinical Trials; Cytokinesis; Development; Dimensions; Embryo; Ensure; Failure; Genetic Materials; Human; Location; Malignant Neoplasms; Mediating; Microtubules; Mitosis; Mitotic; Molecular; Monitor; Mothers; Motor; Mutation; Myosin Type II; Pattern; Phosphotransferases; Positioning Attribute; Process; Proteins; Research; Role; Signal Transduction; Site; Specific qualifier value; Techniques; Testing; Therapeutic; Up-Regulation; Work; anillin; assay development; aurora kinase A; blebbistatin; cancer type; cell cortex; chemotherapeutic agent; chemotherapy; constriction; daughter cell; experimental study; inhibitor; insight; interest; overexpression; therapeutic target; tumorigenesis

PROJECT SUMMARY/ABSTRACT Cytokinesis is the final step in mitosis and facilitates the physical cleavage of one cell into two daughter cells. Animal cells achieve cytokinesis through the constriction of a cortical actomyosin contractile ring that forms around the cell equator between the segregating chromosomes. Failure of cytokinesis can lead to the formation of tetraploid cells that are thought to be an important intermediate in tumorigenesis. To ensure equal partitioning of the genetic material, contractile ring assembly is controlled by the anaphase spindle. The anaphase spindle sends two superimposed signals that pattern the contractility of the overlying cortex; a signal from the central spindle that forms between the separating chromosomes promotes contractility of the equatorial cortex, and a signal from the centrosomal microtubule asters suppresses contractility of the non-equatorial cortex. Recent work in C. elegans in my sponsor lab has implicated the mitotic kinase Aurora A in suppressing the accumulation of contractile ring proteins on the non-equatorial cortex. However, whether this function of Aurora A is conserved in human cells and the identity of the Aurora A target sites that mediate suppression of cortical contractility are currently unknown. In the proposed work, I will determine whether the role of Aurora A in patterning cortical contractility is conserved in human cells, use molecular replacement techniques in C. elegans to identify the targets of Aurora A-mediated suppression of cortical contractility, and determine how Aurora A suppression of contractility is integrated with central spindle promotion of contractility to pattern contractile ring formation. In Aim1, I will develop an assay to monitor the suppression of contractility on the non-equatorial cortex during cytokinesis in human cells and use it to determine if Aurora A has a conserved role in this process and whether ectopic targeting of Aurora A to the plasma membrane can suppress contractility. In Aim 2A, I will capitalize on the advantages the C. elegans embryo offers for molecular replacement experiments to perform an unbiased screen of cytokinesis regulators to identify the Aurora A target sites that mediate the ability of centrosomal asters to suppress cortical contractility and will test identified regulatory sites in human cells for conservation. In Aim 2B I will determine how Aurora A and central spindle-derived signals are integrated to allow for the precise specification of contractile ring location and dimensions. Collectively, this work will provide crucial insight into how Aurora A contributes to patterning cortical contractility in dividing cells. As inhibitors targeting Aurora A are currently in clinical trials as potential chemotherapeutic agents, understanding how the role of Aurora A in regulating contractility is related to its functions in tumorigenesis has the potential to impact therapeutic strategies in cancer.

项目概要/摘要 细胞分裂是有丝分裂的最后一步，有助于将一个细胞物理分裂成两个 子细胞。动物细胞通过皮质肌动球蛋白的收缩实现胞质分裂 在分离染色体之间的细胞赤道周围形成的收缩环。失败 胞质分裂可以导致四倍体细胞的形成，四倍体细胞被认为是重要的中间体 在肿瘤发生中。为了确保遗传物质的平等分配，收缩环组装是 由后期纺锤体控制。后期纺锤体发送两个叠加信号 模拟上层皮质的收缩性；来自中心主轴的信号，形成于 分离染色体促进赤道皮层的收缩性，并且来自赤道皮层的信号 中心体微管紫苑抑制非赤道皮层的收缩性。最近的工作于 我的赞助者实验室中的线虫表明有丝分裂激酶 Aurora A 与抑制 收缩环蛋白在非赤道皮质上的积累。不过这个功能是否 Aurora A 的基因在人类细胞中是保守的，并且 Aurora A 介导的靶位点的身份 皮质收缩性的抑制目前尚不清楚。在拟议的工作中，我将确定 Aurora A 在皮质收缩模式中的作用在人类细胞中是否保守，使用 秀丽隐杆线虫中的分子替代技术，用于识别 Aurora A 介导的靶标 抑制皮质收缩性，并确定 Aurora A 抑制收缩性的效果如何 与中央纺锤体促进收缩性相结合，形成收缩环模式。在目标1中，我 将开发一种检测方法来监测非赤道皮层收缩力的抑制 人类细胞中的胞质分裂，并用它来确定 Aurora A 在此过程中是否具有保守作用，以及 Aurora A 异位靶向质膜是否可以抑制收缩性。在目标 2A 中， 我将利用线虫胚胎为分子替代实验提供的优势 对胞质分裂调节因子进行公正的筛选，以确定介导 Aurora A 的靶位点 中心体紫苑抑制皮质收缩性的能力并将测试已确定的调节位点 在人体细胞中进行保存。在目标 2B 中，我将确定 Aurora A 和中心纺锤体如何衍生 信号被集成以允许收缩环位置和尺寸的精确规范。 总的来说，这项工作将为 Aurora A 如何促进皮质模式提供重要的见解 分裂细胞的收缩力。由于针对 Aurora A 的抑制剂目前正在临床试验中，具有潜力 化疗药物，了解 Aurora A 在调节收缩力方面的作用如何相关 其在肿瘤发生中的功能有可能影响癌症的治疗策略。