Dissecting the Mechanisms of Cell Division

智力优点：有丝分裂纺锤体组装是一个高度复杂和特别精心策划的事件，是细胞分裂所必需的。它依赖于大量的蛋白质复合物，蛋白质-蛋白质相互作用和由酶活性驱动的调节机制。目前的进展，以产生一个全面的地图的途径，是至关重要的主轴组装已阻碍了两个主要的障碍。首先，细胞分裂酶的互补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，并可用于以急性和暂时的方式定义其功能。为了进一步了解纺锤体组装和细胞分裂所需的基本机制，我们正在采取两种互补的多学科方法。首先，我们正在表征20种新的细胞分裂酶，这些酶在我们的蛋白质组学和遗传学筛选中被鉴定为它们的耗竭表型、亚细胞定位和相互作用的伴侣。这些数据将用于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。第二，我们正在测试新型小分子作为化学探针的效用，用于解剖这些关键细胞分裂酶的机制。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。这项研究将促进我们对细胞通路、酶组分以及这些酶促进细胞正常分裂的作用机制的理解。更广泛的影响：这项多学科研究将为来自细胞生物学、化学、生物化学和生物工程等多个领域的高中生、本科生和研究生创造指导、教育和培训机会。特别是将鼓励妇女和在科学领域代表性不足的少数群体参加。学生将通过研讨会和实践培训模块接受先进生物化学研究方法和仪器的指导和培训。这将使学生能够进行独立研究，有助于我们对调节细胞分裂机制的理解，并获得对科学过程的终身赞赏。这项研究将产生大量的数据集和工具，这将有利于科学界，并可用于进一步了解细胞如何分裂。所有研究成果将在期刊上广泛传播，作为补充材料，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邀请演讲中。这项研究将通过激励年轻学生并使他们能够欣赏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.