Centrosomin and Centrosomes in Cell Division

细胞分裂中的中心体蛋白和中心体

基本信息

批准号：
7268755
负责人：
TIMOTHY L MEGRAW
金额：
$ 25.89万
依托单位：
UT SOUTHWESTERN MEDICAL CENTER
依托单位国家：
美国
项目类别：
财政年份：
2003
资助国家：
美国
起止时间：
2003-08-01 至 2009-07-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7268755
关键词：
Amino Acids Aneuploidy Animals Antibodies Binding Biochemical Genetics Biological Assay Cell Cycle Stage Cell Death Cell Extracts Cell Line Cell division Cell model Cell physiology Cells Centrosome Chimeric Proteins Chromosomal Stability Chromosome Segregation Chromosomes Coiled-Coil Domain Complex Cultured Cells Cytogenetics Cytoskeletal Modeling Cytoskeleton Drosophila genus Epitopes Female Genes Genetic Glutathione Goals Growth Homologous Gene Human Immunoprecipitation Incubated Length Leucine Zippers Maps Meiosis Microtubule-Organizing Center Microtubules Mitosis Mitotic Mitotic Spindle Apparatus Mitotic spindle Mutate Normal Cell Oncogenic Organism Pathway interactions Phosphoproteins Phosphorylation Phosphorylation Site Play Process Protein Fragment Protein Phosphatase 2A Regulatory Subunit PR53 Protein phosphatase Proteins Purpose RNA Interference Recruitment Activity Regulation Role Sepharose Site Somatic Cell Structure Vertebrates Yeasts carcinogenesis cell type chromosome movement cofactor daughter cell design in vivo knock-down mutant novel protein function research study yeast two hybrid system

项目摘要

DESCRIPTION (provided by applicant): The equal distribution of chromosomes into daughter cells depends upon the accurate assembly of the mitotic spindle at cell division. The inappropriate segregation of chromosomes leads to aneuploidy, which can result in cell death or carcinogenesis. Proteins that control the cytoskeleton, including those that regulate microtubule dynamics and spindle assembly, can be oncogenic when altered. Our goal is to understand the function of the centrosome in cytoskeletal organization and other cellular processes. To achieve this, we must understand the roles of proteins that function at the centrosome. Our main focus is on Centrosomin (Cnn), a major core component of centrosomes in Drosophila. Cnn is a phosphoprotein with three putative leucine zippers, extensive coiled-coil domains, and two novel motifs that are highly conserved across species. Cnn is required to assemble functional centrosomes at mitosis. In somatic cells that lack Cnn, mitosis occurs efficiently, but by an alternate, anastral pathway. We hypothesize that the anastral pathway for mitosis may be a vestige of an evolutionary precursor to the current centrosomal mode of mitosis that is found in most animal cells. The purpose of this project is to understand the functions of Cnn and centrosomes in cell division, and to define the anastral pathway for mitosis. Three specific aims are designed to accomplish these goals. The first aim is to dissect the functional domains in Cnn protein using in vivo functional and rescue assays, to examine potential human homologs by rescue of Cnn in Drosophila, and to examine the role of phosphorylation in Cnn function. Our second aim is to identify Cnn protein partners using genetic and biochemical approaches, then assay their localization in the cell, their role in centrosome function, and their role in centrosomal localization of Cnn. Our third aim is to define the anastral meiosis-like pathway that is induced in cells that lack Cnn. The mechanisms for spindle assembly and chromosome segregation at mitosis in cells that lack Cnn (and have dysfunctional centrosomes) will be investigated.

描述（由申请人提供）：染色体在子细胞中的相等分布取决于细胞分裂中有丝分裂纺锤体的精确组装。染色体的不当分离导致非整倍性，这可能导致细胞死亡或致癌作用。控制细胞骨架的蛋白质，包括调节微管动力学和纺锤体组件的蛋白质，在改变时可能是致癌性的。我们的目标是了解中心体在细胞骨架组织和其他细胞过程中的功能。为了实现这一目标，我们必须了解在中心体中起作用的蛋白质的作用。我们的主要重点是果蝇中心体的主要核心成分中心素（CNN）。 CNN是一种磷酸蛋白，具有三个推定的亮氨酸拉链，广泛的盘绕螺旋域和两个在物种之间高度保守的新基序。 CNN需要在有丝分裂时组装功能性中心体。在缺乏CNN的体细胞中，有丝分裂有效地发生，但通过替代的斜途径。我们假设有丝分裂的肛门途径可能是大多数动物细胞中发现的当前有丝分裂中心体模式的进化前体的遗迹。该项目的目的是了解CNN和中心体在细胞分裂中的功能，并定义有丝分裂的厌氧途径。旨在实现这些目标的三个特定目标。第一个目的是使用体内功能和救援测定法剖定CNN蛋白中的功能结构域，以检查果蝇中CNN的潜在人类同源物，并检查磷酸化在CNN功能中的作用。我们的第二个目的是使用遗传和生化方法鉴定CNN蛋白质伴侣，然后测定它们在细胞中的定位，其在中心体功能中的作用以及它们在CNN的中心体定位中的作用。我们的第三个目的是定义缺乏CNN的细胞引起的类似减数分裂的途径。将研究缺乏CNN（并且具有功能失调的中心体）细胞中纺锤体组装和染色体分离的机制。