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.