Centrosome Maturation In Vitro

中心体体外成熟

• 批准号: 6829661
• 负责人: ROBERT E PALAZZO
• 金额: $ 27.06万
• 依托单位: RENSSELAER POLYTECHNIC INSTITUTE
• 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 2007-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6829661
• 关键词: Antibodies; Asses; Biochemical; Cell Cycle; Cell Shape; Cell physiology; Cells; Centrosome; Chemotaxis; Code; Development; Enzymes; Funding; Genes; Goals; In Vitro; Maintenance; Methods; Microtubule-Organizing Center; Microtubules; Molecular; Neoplasm Metastasis; Oocytes; Organelles; Phase; Phosphorylation; Property; Protein Kinase; Proteins; Regulation; System; Work; Yeasts; base; cell motility; man; trafficking

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，参与从酵母到人的细胞结构的组织。作为微管组织中心，中心体直接或间接参与了许多基本的细胞过程，包括细胞复制、细胞迁移、细胞器定向运输以及细胞形状和极性的维持。因此，了解中心体的功能将影响我们对癌症、转移、趋化和早期发育的理解。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以及中心体功能的生化调节。利用Spisula solidissima卵母细胞的独特特性，已开发出以下方法：1)在体外诱导细胞周期特异性中心体成熟；2)从卵母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；以及5)分离中心体蛋白编码基因。需要资金继续工作，以确定中心体组装和中心体依赖的微管成核所需的蛋白质。将研究中心体蛋白的磷酸化，以确定控制中心体的蛋白激酶酶，以开始揭示蛋白质磷酸化调节中心体功能的机制。这项提议将完成识别重要中心体蛋白的工作，并开始利用这一独特的体外系统来纯化调节中心体功能的分子。

项目成果

Dynein is required for spindle assembly in cytoplasmic extracts of Spisula solidissima oocytes.

在 Spisula Solidissima 卵母细胞的细胞质提取物中，纺锤体的组装需要动力蛋白。

• DOI: 10.1242/jcs.112.9.1291
• 发表时间: 1999
• 期刊: Journal of cell science
• 影响因子: 4
• 作者: Palazzo,RE;Vaisberg,EA;Weiss,DG;Kuznetsov,SA;Steffen,W
• 通讯作者: Steffen,W

Regulation of M-phase progression in Chaetopterus oocytes by protein kinase C.

蛋白激酶 C 调节毛鳍鱼卵母细胞的 M 期进程。

• DOI: 10.1016/0012-1606(92)90294-q
• 发表时间: 1992
• 期刊: Developmental biology
• 影响因子: 2.7
• 作者: Eckberg,WR;Palazzo,RE
• 通讯作者: Palazzo,RE

Reconstitution of centrosome microtubule nucleation in Spisula.

Spisula 中心体微管成核的重建。

• DOI: 10.1016/s0091-679x(01)67011-0
• 发表时间: 2001
• 期刊: Methods in cell biology
• 作者: Schnackenberg,BJ;Palazzo,RE
• 通讯作者: Palazzo,RE

Using centrosome fragments in the directed assembly of microtubules.

在微管的定向组装中使用中心体片段。

• DOI: 10.1166/jnn.2009.c043
• 发表时间: 2009
• 期刊: Journal of nanoscience and nanotechnology
• 作者: Shang,Wen;Crone,DonnaE;Yang,Hoichang;Dordick,JonathanS;Palazzo,RobertE;Siegel,RichardW
• 通讯作者: Siegel,RichardW

Centrosome maturation.

• DOI: 10.1016/s0070-2153(99)49021-0
• 发表时间: 2000
• 期刊: Current topics in developmental biology
• 作者: R. Palazzo;J. Vogel;B. J. Schnackenberg;Dawn R. Hull;Xingyong Wu