Centrosome Maturation In Vitro

中心体体外成熟

• 批准号: 6434202
• 负责人: ROBERT E PALAZZO
• 金额: $ 26.67万
• 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 2002-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6434202
• 关键词: Bivalvia; cell component structure /function; cell cycle; cell morphology; cell proliferation; cellular polarity; centrosome; complementary DNA; egg /ovum; high performance liquid chromatography; high voltage electron microscopy; intracellular; microtubule associated protein; microtubules; molecular cloning; monoclonal antibody; phosphoproteins; phosphorylation; polymerase chain reaction; protein binding; protein kinase; protein protein interaction; tubulin

Centrosomes are unique subcellular organelles involved in the organization of cytoarchitecture from yeast to man. Ass microtubule organizing centers (MTOCs), centrosomes are directly or indirectly involved in numerous fundamental cell processes including cell replication, cell migration, directed organelle traffic and maintenance of cell shape and polarity. Thus, understanding centrosome function will impact our understanding of cancer, metastasis, chemotaxis and early development. The long term goals of this work are to understand centrosome composition, the molecular basis of microtubule nucleation and the biochemical regulation of centrosome function. Taking advantage of the unique properties of Spisula solidissima oocytes, methods have been developed to: 1) induce cell cycle-specific centrosome maturation in vitro; 2) isolate centrosomes from distinct phases of the oocyte cell cycle, 3) disassemble and reassemble a centrosome's ability to organize microtubules, 4) generate antibodies which specifically recognize centrosome proteins, and 5) isolate genes that code for centrosome proteins. Funds are requested to continue work to identify proteins required for centrosome assembly and centrosome- dependent microtubule nucleation. Centrosome protein phosphorylation will be studied to identify protein kinase enzymes that control centrosomes, to begin to unravel the mechanisms by which protein phosphorylation regulates centrosome function. This proposal will complete work to identify important centrosome proteins, and begin to exploit this unique in vitro system to purify molecules that regulate centrosome function.

中心体是一种独特的亚细胞器，参与从酵母到人类的细胞结构的组织，与微管组织中心（MTOC）一样，直接或间接地参与细胞的许多基本过程，包括细胞复制、细胞迁移、定向细胞器运输以及维持细胞的形状和极性。因此，了解中心体功能将影响我们对癌症，转移，趋化性和早期发展的理解。这项工作的长期目标是了解中心体的组成，微管成核的分子基础和中心体功能的生化调节。利用刺海鞘卵母细胞的独特性质，人们已经发展了以下方法：1）体外诱导细胞周期特异性中心体成熟; 2）从卵母细胞细胞周期的不同阶段分离中心体，3）分解和重组中心体组织微管的能力，4）产生特异性识别中心体蛋白的抗体，和5）分离编码中心体蛋白的基因。要求提供资金，以继续鉴定中心体组装和中心体依赖性微管成核所需的蛋白质。中心体蛋白磷酸化将被研究，以确定控制中心体的蛋白激酶，开始解开蛋白磷酸化调节中心体功能的机制。这项建议将完成工作，以确定重要的中心体蛋白，并开始利用这种独特的体外系统，以纯化分子，调节中心体功能。