Mechanistic Analysis of Cytokinesis in Eukaryotes

真核生物细胞分裂的机制分析

• 批准号: 9119026
• 负责人: Erfei Bi
• 金额: $ 43万
• 依托单位: UNIVERSITY OF PENNSYLVANIA
• 依托单位国家: 美国
• 财政年份: 2015
• 资助国家: 美国
• 起止时间: 2015-08-01 至 2019-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9119026
• 关键词: Actomyosin; Address; Anemia; Aneuploidy; Animals; Architecture; Biological Assay; Biology; C2 Domain; CRISPR/Cas technology; Cell Count; Cells; Chitin Synthase; Complex; Cytokinesis; Data; Defect; Development; Disease; Elements; Enzymes; Eukaryota; Evolution; Experimental Models; Extracellular Matrix; Failure; Filament; Genomic Instability; Health; Homeostasis; In Vitro; Liposomes; Malignant Neoplasms; Mammalian Cell; Mediating; Membrane; Membrane Fusion; Microfilaments; Microscopy; Modeling; Myosin ATPase; Myosin Type II; Neck; Neurons; Organism; Platinum; Play; Ploidies; Prevalence; Process; Protein Isoforms; Proteins; Regulation; Research; Resolution; Role; Saccharomyces cerevisiae; Saccharomycetales; Sarcomeres; Site; Structure; System; Technology; Testing; Tissues; Transglutaminases; Vesicle; Yeasts; base; cell type; constriction; design; genome editing; human disease; in vivo; innovation; innovative technologies; novel; phenomenological models; scaffold; success

 DESCRIPTION (provided by applicant): Cytokinesis is essential for development and survival of all organisms. Defects in cytokinesis cause aneuploidy and genomic instability, and thereby contribute to serious diseases such as cancer, neuronal disorders, and anemia. Thus, mechanistic study of cytokinesis is important not only for understanding the basic principles of a fundamental process but also for designing new strategies to treat human diseases. Cytokinesis in animal and fungal cells requires concerted functions of a contractile actomyosin ring (AMR), targeted vesicle fusion, and localized extracellular matrix (ECM) remodeling. Despite extensive studies of cytokinesis over a century, the basic architecture of the AMR remains unknown in any system. It is also unclear how myosin-II localization and filament assembly are regulated during cytokinesis. Increasing evidence suggests that ECM remodeling is critical for cytokinesis not only for yeast cells but also for animal cells. However, the underlying mechanisms remain poorly understood. We propose to address these key questions in cytokinesis using both budding yeast and mammalian cells as our experimental models. In Aim 1, we will determine the AMR structure in budding yeast and then develop a quantitative model for it. This model will open new avenues to address broad questions in cytokinesis, e.g. those regarding the mechanism of ring constriction as well as those concerning the assembly, regulation, and function of the ring. We will also examine the AMR in mammalian cells to establish the degree of conservation in this core cytokinetic structure. In Aim 2, we will test our hypothesis that IQGAP functions as a dual regulator of myosin localization and filament assembly during cytokinesis in both budding yeast and mammalian cells. In Aim 3, we will test our hypothesis that the AMR-associated protein Inn1 interacts with SNAREs via its C2 domain to facilitate vesicle fusion at the division site, thereby increasing the local concentration of the cargo enzyme Chs2 (chitin synthase-II), which is subsequently activated by Cyk3 via its transglutaminase-like domain to promote septum formation (equivalent of ECM remodeling in animal cells) during cytokinesis. The proposed research is innovative, as diverse and cutting-edge technologies will be applied to generate new information and concepts regarding the core mechanisms of cytokinesis.

 描述（适用提供）：细胞因子对于所有组织的发展和生存至关重要。细胞因子的缺陷会导致非整倍性和基因组不稳定性，从而导致严重的疾病，例如癌症，神经元疾病和贫血。这是细胞因子的机械研究不仅对于理解基本过程的基本原理，而且对于设计治疗人类疾病的新策略而言很重要。动物和真菌细胞中的细胞因子需要收缩肌动蛋白环（AMR），靶向场地融合和局部细胞外基质（ECM）重塑的协同功能。尽管对一个多世纪以来的细胞因子进行了广泛的研究，但在任何系统中，AMR的基本结构仍然尚不清楚。还不清楚在细胞因子过程中如何调节肌球蛋白-II定位和细丝组件。越来越多的证据表明，ECM重塑对细胞因子不仅对酵母细胞而言至关重要，而且对动物细胞也至关重要。但是，基本机制仍然知之甚少。我们建议使用萌芽的酵母和哺乳动物细胞作为我们的实验模型解决这些关键问题。在AIM 1中，我们将确定萌芽酵母中的AMR结构，然后为其开发定量模型。该模型将开辟新的途径，以解决细胞动力学中的广泛问题，例如与环的机理以及有关环的组装，调节和功能的机制。我们还将检查哺乳动物细胞中的AMR，以建立这种核心细胞动力学结构中的保护程度。在AIM 2中，我们将检验我们的假设，即IQGAP在萌芽的酵母菌和哺乳动物细胞中都可以作为肌球蛋白定位和细丝组件的双重调节剂。 In Aim 3, we will test our hypothesis that the AMR-associated protein Inn1 interacts with SNAREs via its C2 domain to facilitate vegetables fusion at the division site, thereby increasing the local concentration of the cargo enzyme Chs2 (chitin synthase-II), which is subsequently activated by Cyk3 via its transglutaminase-like domain to promote septum formation (equivalent of ECM在动物细胞中重塑）在细胞因子过程中。拟议的研究具有创新性，因为潜水员和尖端技术将用于生成有关细胞因子核心机制的新信息和概念。