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Probing the spatiotemporal regulation of cell division

探讨细胞分裂的时空调控

基本信息

批准号：
10456882
负责人：
JULIE C CANMAN
金额：
$ 31.65万
依托单位：
COLUMBIA UNIVERSITY HEALTH SCIENCES
依托单位国家：
美国
项目类别：
财政年份：
2019
资助国家：
美国
起止时间：
2019-09-17 至 2024-07-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10456882
关键词：
Actins Actomyosin Address Adult Animals Anterior Biological Process Caenorhabditis elegans Cell Cycle Cell Cycle Regulation Cell Nucleus Cell division Cells Chromatin Collection Complex Cytokinesis Data Defect Development Embryo Ensure Foundations Genetic Gonadal structure Heating Hematological Disease Human Human Pathology In Situ Individual Inherited Kinetics Lasers Lead Light Location Malignant Neoplasms Manuscripts Mediating Methods Microscope Microscopy Microtubules Modeling Molecular Morphogenesis Myosin ATPase Myosin Type II Nature Neurologic Phenotype Population Positioning Attribute Procedures Protein Inhibition Proteins Publishing Regulation Relaxation Role Signal Transduction Site Somatic Cell Sterility Subcutaneous Tissue System Technology Temperature Testing Time Tissues Vulva aurora B kinase base cell cortex cell type comparative constriction daughter cell embryo cell embryo membrane experimental study genetic approach human disease mutant nervous system disorder novel optogenetics precursor cell predictive modeling protein function spatiotemporal stem cells subcellular targeting temperature sensitive mutant temporal measurement tool

项目摘要

PROJECT SUMMARY In animal cells, cytokinesis is driven by constriction of an actomyosin contractile ring, which is positioned and controlled by signaling from spindle microtubules. Cytokinesis requires a high degree of spatial and temporal molecular regulation to ensure each daughter cell inherits a single nucleus. Although the essential molecular players required for cytokinesis are known, many cytokinesis proteins localize dynamically to multiple subcellular niches throughout cell division, potentially allowing multiple subcellular functions and making these proteins difficult study using traditional genetic approaches. Optogenetics enable spatiotemporal studies by locally targeting light to control protein function in specific subcellular regions. For this reason, we developed FLIRT (Fast Local Infrared Thermogenetics), which uses an infrared (IR) laser to locally heat and thus locally inactivate genetically-encoded fast-acting temperature sensitive (ts) mutant proteins with high spatiotemporal precision. FLIRT is also reversible: the IR laser can be turned off at any point to stop local heating and allow for protein reactivation. Furthermore, using FLIRT, non-ts-mutants (wildtype) can be used as controls for any laser-induced damage induced by a given FLIRT procedure. In preliminary data using C. elegans embryos, we calibrated the temperature induction achieved using FLIRT, validated the use of FLIRT on the subcellular level in both the 1-cell embryo and the 16-cell embryo, and demonstrated that FLIRT can inhibit other cell biological processes such as cell fate signaling in multicellular embryos and membrane partitioning in the adult gonad. Having laid the foundation for further studies, we now propose experiments to define the spatiotemporal regulation of actomyosin contractility and spindle microtubule-associated signaling complexes during cytokinesis and address longstanding questions in the field, such as the relative contributions of equatorial vs. polar actomyosin contractility during cell division. FLIRT experiments will also be conducted on cells in C. elegans early embryos and in somatic cells with multicellular developing worm tissue. These experiments will define the precise spatiotemporal regulation of key players in cytokinesis, test specific hypotheses regarding their mechanisms of action, and test the universality of rules governing cytokinesis, whether the same principles that apply to the early embryo also apply to somatic cells within a multicellular context.

项目总结