Molecular Genetic Analysis of Regulatory Networks Governing Cytokinesis

细胞分裂调控网络的分子遗传学分析

• 批准号: RGPIN-2014-03944
• 负责人: Karagiannis, Jim
• 金额: $ 2.55万
• 依托单位: University of Western Ontario
• 依托单位国家: 加拿大
• 项目类别: Discovery Grants Program - Individual
• 财政年份: 2018
• 资助国家: 加拿大
• 起止时间: 2018-01-01 至 2019-12-31
• 项目状态: 已结题
• 来源: https://www.nserc-crsng.gc.ca/ase-oro/Details-Detailles_eng.asp?id=655086
• 关键词: Molecular; Genetic; Analysis; Regulatory; Networks

Research within the lab focuses on the regulatory networks required to bring about the successful completion of cytokinesis in the model eukaryote, Schizosaccharomyces pombe. While previous work in the field has centered on the cytoskeletal structures and forces required to bring about cell cleavage, our goals are to better understand the genetic regulatory networks required for cytokinesis to occur faithfully, at the proper location, and at the proper time during the cell cycle.* *The foundation of the current proposal stems from recent work supporting the existence of a cytokinesis "monitoring system". Upon perturbation of the cell division machinery, this system has the capacity to both delay cell cycle progression and stabilize the cytokinetic actomyosin ring. The biological importance of such mechanisms have been substantiated in recent years by the demonstration that tetraploid intermediates (produced upon cytokinesis failure) show a markedly increased potential for tumourigenesis. Thus, an understanding of these networks has the potential to provide insight into the mechanisms important for maintaining genomic integrity in eukaryotes.**The current proposal revolves around the molecular and genetic analysis of several previously uncharacterized regulatory networks required for the proper function and/or regulation of the cytokinesis monitoring system. These pathways were identified through the execution of a genome-wide screen that assayed fission yeast gene deletion mutants for hypersensitivity to Latrunculin A (an actin depolymerizing drug used as a tool to perturb the cell division machinery). As expected this screen identified many genes with known roles in the regulation of cytoskeletal dynamics and/or cytokinesis. In addition, many other unexpected "hits", identifying genes affecting biological processes such as histone acetylation/de-acetylation and transcriptional control were also identified.**Using the lab's established skill in molecular biology, genetic analysis, and live-cell imaging, our objectives are to characterize the identified regulatory complexes in order to provide a comprehensive, genome-wide picture of the genetic regulatory wiring required by a model eukaryotic cell to faithfully and reliably execute cytokinesis.

实验室内的研究集中在成功完成真核生物模式裂殖酵母胞质分裂所需的调控网络上。虽然该领域以前的工作主要集中在细胞骨架结构和导致细胞分裂所需的力量上，但我们的目标是更好地了解细胞质分裂在细胞周期中忠实地、在适当的位置和适当的时间发生所需的遗传调控网络。**当前提议的基础源于最近支持细胞质分裂“监测系统”的存在的工作。在细胞分裂机制的干扰下，该系统既具有延缓细胞周期进程的能力，又具有稳定细胞动力肌球蛋白环的能力。近年来，四倍体中间体(在胞质分裂失败时产生)显示出显着增加的肿瘤发生潜力，证实了这种机制的生物学重要性。因此，对这些网络的了解有可能为真核生物维持基因组完整性的重要机制提供洞察。**当前的提议围绕着对几个以前未描述的调控网络的分子和遗传分析，这些调控网络是胞质分裂监测系统正常功能和/或调节所必需的。这些途径是通过执行全基因组筛查来确定的，该筛查测试了分裂酵母基因缺失突变体对Latrunculin A(一种肌动蛋白解聚药物，用作扰乱细胞分裂机制的工具)的超敏反应。正如预期的那样，这一筛选发现了许多在细胞骨架动力学和/或胞质分裂的调节中具有已知作用的基因。此外，还识别了许多其他意想不到的“成功”，识别影响生物过程的基因，如组蛋白乙酰化/去乙酰化和转录控制。**利用实验室在分子生物学、遗传分析和活细胞成像方面的成熟技能，我们的目标是表征已识别的调控复合体，以便提供模型真核细胞忠实和可靠地执行胞质分裂所需的基因调控连线的全面、全基因组的图景。