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-2019-05380
• 负责人: Karagiannis, Jim
• 金额: $ 3.06万
• 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: 2021
• 资助国家: 加拿大
• 起止时间: 2021-01-01 至 2022-12-31
• 项目状态: 已结题
• 来源: https://www.nserc-crsng.gc.ca/ase-oro/Details-Detailles_eng.asp?id=738236
• 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 cleavage in cells grown under ideal conditions, our goals are to better understand the genetic regulatory networks required to prevent cytokinesis failure when cells are subject to stresses that impede the cell division process. 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 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 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 recent 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 roles in the regulation of cytoskeletal dynamics and/or cytokinesis. In addition, many other unexpected "hits", identifying genes affecting biological processes such as stress-dependent signaling, histone acetylation/de-acetylation, and transcriptional control were also identified. Using the lab's expertise in molecular biology, genetic analysis, and fluorescence microscopy, our objectives are to characterize the identified regulatory complexes in order to provide a comprehensive, genome-wide picture of the genetic regulatory wiring and molecular mechanisms required by a model eukaryotic cell to faithfully and reliably execute cytokinesis.

实验室的研究重点是在真核生物裂糖酵母（Schizosaccharomyces pombe）中成功完成细胞分裂所需的调控网络。虽然该领域以前的工作集中在理想条件下生长的细胞中产生分裂所需的细胞骨架结构和力，但我们的目标是更好地理解当细胞受到阻碍细胞分裂过程的压力时防止细胞分裂失败所需的遗传调控网络。当前建议的基础源于最近支持细胞分裂“监测系统”存在的工作。当细胞分裂机制受到干扰时，该系统具有延迟细胞周期进程和稳定细胞动力学肌动球蛋白环的能力。这种机制的生物学重要性已被证实，证明四倍体中间体（在细胞分裂失败时产生）显着增加了肿瘤发生的潜力。因此，对这些网络的理解有可能提供对维持真核生物基因组完整性的重要机制的见解。目前的建议围绕着先前未表征的调节网络的分子和遗传分析，这些网络是细胞分裂监测系统的正常功能和/或调节所必需的。这些途径是通过最近的全基因组筛选确定的，该筛选分析了裂变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，该筛选确定了许多在细胞骨架动力学和/或细胞分裂调节中起作用的基因。此外，还发现了许多其他意想不到的“命中”，确定了影响生物过程的基因，如应激依赖性信号，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、遗传分析和荧光显微镜方面的专业知识，我们的目标是表征已识别的调控复合物，以便提供一个完整的、全基因组的遗传调控线路和模型真核细胞所需的分子机制，以忠实可靠地执行细胞分裂。