Mechanistic Analysis of Cytokinesis in Eukaryotes

真核生物细胞分裂的机制分析

基本信息

批准号：
10451747
负责人：
Erfei Bi
金额：
$ 44.78万
依托单位：
UNIVERSITY OF PENNSYLVANIA
依托单位国家：
美国
项目类别：
财政年份：
2015
资助国家：
美国
起止时间：
2015-08-01 至 2024-07-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10451747
关键词：
Actins Actomyosin Address Affect Anemia Aneuploidy Animal Model Animals Architecture Biochemical Genetics Biological Models Biology Biosensor C2 Domain Cell model Cells Clustered Regularly Interspaced Short Palindromic Repeats Complex Coupled Cytokinesis Defect Deposition Development Disease Electron Microscopy Enzymatic Biochemistry Enzymes Eukaryota Excision Exocytosis Extracellular Matrix Failure Family Filament Fission Yeast Genomic Instability Goals Guanine Nucleotide Exchange Factors Hela Cells Human Image Label Life Malignant Neoplasms Mammalian Cell Mediating Microfilaments Microscopy Molecular Monitor Myosin ATPase Myosin Type II Neurons Optics Organism Phosphorylation Platinum Positioning Attribute Process Production Protein Isoforms Proteins Recombinants Regulation Research Resolution Role Saccharomyces cerevisiae Saccharomycetales Site System Tail Testing Trans-Activators Transglutaminases Vesicle Yeasts base design glycosyltransferase human disease imaging modality in vivo innovation member monomer non-muscle myosin novel protein complex rab GTP-Binding Proteins reconstruction spatiotemporal vesicle transport

项目摘要

Project Summary/Abstract: Cytokinesis is essential for development and survival of all organisms. Defects in cytokinesis cause aneuploidy and genomic instability, thereby contributing 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 spatiotemporally coordinated functions of a contractile actomyosin ring (AMR), targeted vesicle fusion, and localized extracellular matrix (ECM) remodeling. It is much more complex than previously appreciated. In this application, we will address three major unanswered questions regarding this fundamental process using both budding yeast and mammalian cell models, with the goal of dissecting deep mechanisms in yeast and exploring evolutionary conservation in mammalian cells. In Aim 1, we will determine the architecture of the AMR. Specifically, we will examine how myosin-II and its associated proteins such as actin and IQGAP are organized in the contractile ring from cells synchronized at cytokinesis using platinum-replica electron microscopy (PREM) coupled with immuno-gold labeling as well as super-resolution stochastic optical reconstruction microscopy (STORM). In Aim 2, we will test our hypothesis that myosin filament assembly is regulated by heavy chain phosphorylation as well as by trans-acting factors such as IQGAP using biochemical, genetic, quantitative live imaging, and other cutting- edge imaging methods as described above. In Aim 3, we will determine how the AMR guides exocytosis and ECM remodeling at the division site. Specifically, we will test our hypothesis that the tail of the yeast myosin-II positions and unloads vesicles from the transport machinery at the division site by interacting with the vesicle-associated guanine-nucleotide-exchange factor (GEF), Sec2, for the Rab GTPase Sec4. Then, the myosin-associated protein complex (Inn1, Hof1, and Cyk3) promotes vesicle fusion via the C2 domain of Inn1, and activates the cargo enzyme Chs2, a member of the glycosyltransferase family 2, for ECM remodeling (i.e. septum formation in yeast) via the transglutaminase-like domain of Cyk3. The discovery (Aim 1) and hypothesis-driven (Aims 2 and 3) research is expected to generate novel concepts and mechanisms of cytokinesis that are beyond specific model organisms.

项目摘要/摘要：细胞因子对于所有生物的发育和生存至关重要。细胞因子的缺陷原因非整倍性和基因组不稳定性，从而导致严重疾病，例如癌症，神经元疾病和贫血。因此，细胞因子的机械研究不仅对于理解基本过程的基本原则，也用于设计治疗人类疾病的新策略。动物和真菌细胞中的细胞因子需要收缩的时空协调功能肌球蛋白环（AMR），靶向囊泡融合和局部的细胞外基质（ECM）重塑。这是比以前所欣赏的要复杂得多。在此应用程序中，我们将解决三个主要使用萌芽酵母和哺乳动物细胞的有关此基本过程的未解决问题模型，目的是剖析酵母中的深层机制并探索进化保存哺乳动物细胞。在AIM 1中，我们将确定AMR的架构。具体来说，我们将研究如何肌球蛋白-II及其相关蛋白（如肌动蛋白和IQGAP）在收缩环中从细胞中组织使用铂 - 替代电子显微镜（PER）与免疫金同步在细胞因子上同步标记以及超分辨率随机光学重建显微镜（Storm）。在AIM 2中，我们将检验我们的假设，即肌球蛋白丝组件受重链磷酸化的调节以及使用生化，遗传，定量实时成像和其他剪切的跨作用因子，例如IQGAP 边缘成像方法如上所述。在AIM 3中，我们将确定AMR指南如何胞吐作用和ECM改建在部门地点。具体来说，我们将测试酵母尾的假设通过与 Rab GTPase Sec4的囊泡相关的鸟嘌呤核苷酸交换因子（GEF），SEC2。然后，与肌球蛋白相关的蛋白质复合物（INN1，HOF1和CYK3）通过C2域促进囊泡融合 INN1并激活ECM的糖基转移酶家族的货物CHS2（糖基转移酶家族的成员）通过cyk3的跨谷氨酰胺酶样结构域的重塑（即酵母中的隔膜形成）。发现（目标1）和假设驱动的（目标2和3）的研究有望产生新颖超出特定模型生物的细胞因子的概念和机制。

项目成果

期刊论文数量（15）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Defining Functions of Mannoproteins in Saccharomyces cerevisiae by High-Dimensional Morphological Phenotyping.

DOI：
10.3390/jof7090769
发表时间：
2021-09-17
期刊：
Journal of fungi (Basel, Switzerland)
影响因子：
0
作者：
Ghanegolmohammadi F;Okada H;Liu Y;Itto-Nakama K;Ohnuki S;Savchenko A;Bi E;Yoshida S;Ohya Y
通讯作者：
Ohya Y

Unraveling the mechanisms and evolution of a two-domain module in IQGAP proteins for controlling eukaryotic cytokinesis.