CENTROSOME MATURATION IN VITRO

体外中心体成熟

• 批准号: 2181914
• 负责人: ROBERT E PALAZZO
• 金额: $ 16.95万
• 依托单位: UNIVERSITY OF KANSAS LAWRENCE
• 依托单位国家: 美国
• 财政年份: 1992
• 资助国家: 美国
• 起止时间: 1992-08-01 至 1995-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2181914
• 关键词: Bivalvia; DNA replication; Saccharomyces; alternatives to animals in research; cell free system; cell growth regulation; centrosome; egg /ovum; fluorescence microscopy; meiosis; microtubules; monoclonal antibody; protein structure function; saltwater environment; sea urchins; stainings; tubulin

Centrosome are involved in the organization of cytoarchitecture in cells from yeast to man. As organizers of microtubules (microtubule organizing centers, MTOCs), centrosomes are involved in numerous fundamental cell processes including cell migration and cell division. Thus, and understanding of the composition and biochemical regulation of centrosomes will have an impact on understanding cancer, metastasis, chemotaxis, and aspects of early development. Methods have been developed to take advantage of the unique characteristics of clam oocytes which allow the purification of sufficient quantities of centrosomes for preparative and analytical biochemistry. The experiments outlined in this proposal will define the protein composition of centrosomes and determine if compositional changes are a function of the meiotic cell cycle. Functional in vitro assays coupled with video- and electron-microscopy will be used to quantitate centriole content and the ability of purified centrosomes to direct microtubule polymerization. Monoclonal antibodies will be generated for the identification and purification of molecules (MTOCs) which direct microtubule organization. Finally, key experiments will be carried out to determine if centrosomes can be induced to mature in vitro, as indicated by changes in their ability to form and replicate centrioles or their ability to organize microtubules. These studies will open the way for classic biochemical approaches designed to purify molecules which regulate centrosomes during the cell cycle.

中心体参与细胞内细胞结构的组织 从酵母到人。 作为微管的组织者（微管组织 中心，MTOC），中心体参与许多基本细胞 过程包括细胞迁移和细胞分裂。 因此，和 了解中心体的组成和生化调节 将对理解癌症、转移、趋化性和 早期发展的各个方面。 已经开发出方法来采取 利用蛤卵母细胞的独特特性， 纯化足够量的中心体用于制备和 分析生物化学。 本提案中概述的实验将 定义中心体的蛋白质组成并确定是否 成分变化是减数分裂细胞周期的函数。 功能性 体外测定与视频和电子显微镜相结合将用于 定量中心粒含量和纯化中心体的能力 直接微管聚合。 将产生单克隆抗体 用于分子 (MTOC) 的鉴定和纯化，这些分子直接 微管组织。 最后，将进行关键实验 确定是否可以在体外诱导中心体成熟，如 它们形成和复制中心粒的能力或其能力发生变化 组织微管。 这些研究将为经典开辟道路 旨在纯化调节分子的生化方法 细胞周期中的中心体。