FORMATION, INTERACTION & FUNCTION OF SPINDLE COMPONENTS

这项研究的广泛目标是确定中心体和 动点起作用并相互作用，形成有丝分裂纺锤体 脊椎动物，染色体运动的作用力是如何产生的 以及动物细胞中有丝分裂的进展是如何被控制的。 目标1是使用3-D EM断层扫描和免疫金方法来确定 SPICEN 300相对于含有伽马微管蛋白的环分布 在孤立的但有功能的中心体中；目标2是确定极点 在昆虫精母细胞纺锤体中存在射出力，并测试 染色体在染色体上达到的最终位置的假说 纺锤体，就在后期开始之前，由非 由相反的主轴极产生的动粒主轴MTS；目标#3是 评估一个动点拉动另一个动点的假设 在染色体运动期间推动到纺锤体赤道(即，汇聚)， 并确定作为会议基础的结构事件 只包含一个着丝粒的染色体。目标4是评估 国会染色体运动方向的假说已确定 通过它的姐妹运动芯片上的MT数量，并完成我们的3-D 脊椎动物附着在MTS上的动点结构的EM分析； 目标5是测试一种假说，即靶点为抑制因子 由未附着的动点所产生的后期开始与 纺锤体，并确定脊椎动物细胞是否也具有其他检查点 它调节有丝分裂的退出；目标#6检验了这样的假设：mt. 与动粒日冕相关的负端马达，其功能 在染色体附着的过程中，也在后期发挥作用；目标7是 来检验中心体的作用力机制 分裂后期的分离在整个有丝分裂过程中是活跃的。《知识》 从这些研究中获得的信息需要更好地理解 各种出生缺陷综合征和癌症，设计新的治疗方法 控制细胞增殖的策略，以及治疗 其他疾病状态包括转移、关节炎和阿尔茨海默氏症。

The broad objectives of this research are to determine how centrosomes and kinetochores function and interact to form the mitotic spindle in vertebrates, how the forces for chromosome motion are generated and regulated, and how progress through mitosis is controlled in animal cells. Aim #1 is to use 3-D EM tomography and immunogold methods to determine how spicen 300 is distributed relative to the gamma-tubulin-containing rings in isolated but functional centrosomes; Aim #2 is to determine if polar ejection forces are present in insect spermatocyte spindles, and to test the hypothesis that the final position a chromosome achieves on the spindle, just before anaphase onset, is determined by the number of non- kinetochore spindle Mts generated by the opposing spindle poles; Aim #3 is to evaluate the hypothesis that one kinetochore pulls while the other pushes during chromosome motion to the spindle equator (i.e., congression), and to determine the structural events that underlie congression of chromosomes containing only one kinetochore. Aim #4 is to evaluate the hypothesis at the direction a congressing chromosome moves is determined by the number of Mts on its sister kinetochores, and to complete our 3-D EM analyses on the structure of vertebrate kinetochores attached to Mts; Aim #5 is to test the hypothesis that the target for the inhibitor of anaphase onset produced by unattached kinetochores is associated with the spindle, and to determine if vertebrate cells also possess other checkpoint that regulate exit from mitosis; Aim #6 tests the hypothesis that the Mt minus-end motors associated with the kinetochore corona, which function during chromosome attachment, also function during anaphase; and Aim 7 is to test the hypothesis that the force-producing mechanism for centrosome separation during anaphase is active throughout mitosis. The knowledge obtained from these studies is needed to better understand the etiology of various birth defect syndromes and cancers, to design new therapeutic strategies for the control of cell proliferation, and for the treatment of other disease states including metastasis, arthritis and Alzheimer's.