Investigating the Cell Division Machinery

摘要 细胞分裂是一组复杂的、特别精心策划的事件，最终导致 姐妹染色单体分成两个细胞。它依赖于大量的蛋白质复合体，蛋白质-蛋白质相互作用，以及 翻译后修饰酶活性驱动的调节机制。细胞调节失调 分裂可导致不受限制的或有缺陷的细胞分裂，从而促进染色体的不稳定和 非整倍体，与许多类型的癌症有关。通过蛋白质组和基因筛查，我们 发现了对细胞分裂的保真度至关重要的新蛋白质。我们已经应用了多学科 利用细胞生物学、分子生物学、计算生物学和生物化学来分析 这些蛋白质的功能。这些方法使我们能够定义许多新蛋白质的功能 在关键细胞分裂事件中的关键作用，包括中心体稳态、有丝分裂微管纺锤体 组装、纺锤体组装检查点功能和胞质分裂。此外，我们还使用了化学物质 定义新的细胞分裂抑制剂及其作用机制的生物学方法，我们已经使用过 作为剖析细胞分裂机制和癌症发展的分子探针 治疗学。在这项建议中，我们建议推进我们对蛋白质及其蛋白质谱系的理解 对细胞分裂很重要的机制。这将包括对新奇和糟糕的- 参与中心体动态平衡和有丝分裂纺锤体的微管相关蛋白 组件，以及有助于主轴组件检查点保真度的新型组件。总体而言， 拟议的研究将增加我们对细胞分裂酶谱系的了解，它们 在关键细胞分裂事件中的作用，它们是如何被调节的，以及它们如何相互协调以 确保适当的细胞分裂。这些研究也将促进我们对细胞分裂机制的理解 在人类发育和增殖性疾病中是不受调控的。

SUMMARY Cell division is a complex and specifically orchestrated set of events that culminates in the equal segregation of sister chromatids into two cells. It relies on a multitude of protein complexes, protein-protein interactions, and regulatory mechanisms driven by the activities of posttranslational modification enzymes. Misregulation of cell division can lead to unrestricted or defective cell divisions that can promote chromosomal instability and aneuploidy, which are associated with many types of cancers. Through proteomic and genetic screens we have uncovered novel proteins that are critical to the fidelity of cell division. We have applied multidisciplinary approaches that utilize cell biology, molecular biology, computational biology and biochemistry to analyze the function of these proteins. These approaches have allowed us to define the function of many new proteins with critical roles in key cell division events including centrosome homeostasis, mitotic microtubule spindle assembly, spindle assembly checkpoint function and cytokinesis. Furthermore, we have applied chemical biology approaches to define novel cell division inhibitors and their mechanisms of action, which we have used as molecular probes for dissecting the mechansims of cell division and for the development of cancer therapeutics. In this proposal, we propose to advance our understanding of the repertoire of proteins and their mechanisms that are important to cell division. This will include functional analysis of novel and poorly- characterized microtubule-associated proteins involved in centrosome homeostasis and mitotic spindle assembly, and novel components that contribute to the fidelity of the spindle assembly checkpoint. Overall, the proposed research studies will increase our understanding of the repertoire of cell division enzymes, their function whithin critical cell division events, how they are regulated, and how they coordinate with each other to ensure proper cell division. These studies will also advance our understanding of the cell division mechanisms that are dysregulated in human developmental and proliferative diseases.