Centrosomin and Centrosomes in Cell Division

细胞分裂中的中心体蛋白和中心体

• 批准号: 7101773
• 负责人: TIMOTHY L MEGRAW
• 金额: $ 26.66万
• 依托单位: UT SOUTHWESTERN MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 2003
• 资助国家: 美国
• 起止时间: 2003-08-01 至 2008-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7101773
• 关键词: SDS polyacrylamide gel electrophoresis; cell cycle; centrosome; chromosome movement; immunoprecipitation; mass spectrometry; mitotic spindle apparatus; phosphoproteins; phosphorylation; protein structure function; tissue /cell culture; yeast two hybrid system

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均匀分布取决于细胞分裂时有丝分裂纺锤体的准确组装。染色体的不适当分离会导致非整倍性，从而导致细胞死亡或致癌。控制细胞骨架的蛋白质，包括那些调节微管动力学和纺锤体组装的蛋白质，在发生改变时可能会致癌。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发挥作用的蛋白质的作用。我们的主要关注点是中心体蛋白 (Cnn)，它是果蝇中心体的主要核心成分。 Cnn 是一种磷蛋白，具有三个假定的亮氨酸拉链、广泛的卷曲螺旋结构域和两个在物种间高度保守的新颖基序。有丝分裂时需要 Cnn 来组装功能性中心体。在缺乏 Cnn 的体细胞中，有丝分裂有效地发生，但通过替代的星状途径。我们假设有丝分裂的星状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，并定义有丝分裂的星状途径。为了实现这些目标，我们设计了三个具体目标。第一个目标是使用体内功能和救援分析来剖析 Cnn 蛋白的功能域，通过果蝇中 Cnn 的救援来检查潜在的人类同源物，并检查磷酸化在 Cnn 功能中的作用。我们的第二个目标是使用遗传和生化方法识别 Cnn 蛋白伴侣，然后分析它们在细胞中的定位、它们在中心体功能中的作用以及它们在 Cnn 中心体定位中的作用。我们的第三个目标是定义在缺乏 Cnn 的细胞中诱导的类星体减数分裂途径。将研究缺乏 Cnn（且中心体功能失调）的细胞在有丝分裂时纺锤体组装和染色体分离的机制。