Dissecting the Mechanisms of Cell Division

智力优势：有丝分裂纺锤体组装是一个高度复杂和特别精心编排的事件，是细胞分裂所必需的。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由酶活性驱动的调节机制。目前在生成对主轴装配至关重要的路径的综合地图方面的进展受到两个主要障碍的阻碍。首先，细胞分裂酶的组成以及它们如何相互协调以形成有丝分裂纺锤体在很大程度上是未知的。其次，只有有限数量的化学探针可以抑制这些酶的功能，并可以用来以急性和暂时的方式定义它们的功能。为了促进我们对纺锤体组装和细胞分裂所需的基本机制的理解，我们采取了两种互补的多学科方法。首先，我们在蛋白质组和遗传学筛选中确定了20种新的细胞分裂酶的特性，包括它们的耗竭表型、亚细胞定位和相互作用伙伴。这些数据将被用来构建细胞分裂酶相互作用网络，使有丝分裂酶作为一个生物系统在时间和空间上的研究成为可能。其次，我们正在测试新型小分子作为化学探针的效用，以剖析这些关键的细胞分裂酶的机制。这项研究需要制定新的研究计划，建立跨学科合作，使用先进的技术工具和仪器，以及整合来自不同来源的数据。这项研究将增进我们对这些酶促进细胞正常分裂的细胞途径、酶成分和作用机制的理解。广泛的影响：这项多学科的研究将为包括细胞生物学、化学、生物化学和生物工程在内的多个领域的高中、本科生和研究生创造指导、教育和培训的机会。特别是，将鼓励妇女和科学界代表性不足的少数群体参与。学生将通过研讨会和动手培训模块接受先进的生化研究方法和仪器的指导和培训。这将使学生能够进行独立的研究，有助于促进我们对调节细胞分裂的机制的理解，并获得对科学过程的终身欣赏。这项研究将产生大量的数据集和工具，这将使科学界受益，并可用于进一步了解细胞是如何分裂的。所有研究成果将作为补充材料在期刊上广泛传播，在我们的实验室网站上，在研究会议/会议上，以及在学院和大学的特邀讲座上。这项研究将通过激励年轻学生并使他们认识到科学及其对我们社会进步的重要性而造福社会。它还将指导和培训下一代高技能和多样化的科学家。

Intellectual merit: Mitotic spindle assembly is a highly complex and specifically orchestrated event that is necessary for cell division. It relies on a multitude of protein complexes, protein-protein interactions, and regulatory mechanisms driven by enzymatic activities. Current progress to generating a comprehensive map of the pathways that are critical for spindle assembly has been hampered by two major roadblocks. First, the complement of cell division enzymes and how they coordinate with each other to form the mitotic spindle is largely unknown. Second, there are only a limited number of chemical probes that can inhibit the function of these enzymes and can be used to define their function in an acute and temporal manner. To advance our understanding of the fundamental mechanisms required for spindle assembly and cell division, we are taking two complementary multidisciplinary approaches. First, we are characterizing 20 novel cell division enzymes identified in our proteomic and genetic screens with respect to their depletion phenotype, subcellular localization, and interacting partners. These data will be used to construct a cell division enzyme interaction network that will enable the study of mitotic enzymes as a biological system in time and space. Second, we are testing the utility of novel small molecules as chemical probes for dissecting the mechanisms of these critical cell division enzymes. This research entails the development of new research schemes, the establishment of interdisciplinary collaborations, the use of advanced technological tools and instrumentation, and the integration of data from diverse sources. This research will advance our understanding of the cellular pathways, enzyme components, and the mechanism of action of these enzymes for promoting proper cell division.Broader impacts: This multidisciplinary research will create mentoring, educational, and training opportunities for high school, undergraduate and graduate students from multiple fields including cell biology, chemistry, biochemistry and bioengineering. In particular women and underrepresented minorities in the sciences will be encouraged to participate. Students will receive instruction and training in advanced biochemical research methods and instrumentation through seminars and hands on training modules. This will enable students to conduct independent research, contribute to the advancement of our understanding of the mechanisms regulating cell division and to gain a lifelong appreciation of the scientific process. This research will generate large data sets and tools, which will benefit the scientific community and can be used to further understand how cells divide. All research results will be disseminated broadly in journals, as supplementary material, on our labs website, at research meetings/conferences, and at invited talks at colleges and universities. This research will benefit society by inspiring young students and enabling them to appreciate science and its importance for the advancement of our society. It will also mentor and train the next generation of highly skilled and diverse scientists.