ANALYSIS OF PARTIALLY RECONSTITUTED KINETOCHORE

这个子项目是许多研究子项目中利用 资源由NIH/NCRR资助的中心拨款提供。子项目和 调查员(PI)可能从NIH的另一个来源获得了主要资金， 并因此可以在其他清晰的条目中表示。列出的机构是 该中心不一定是调查人员的机构。 所有细胞的繁殖都需要准确的染色体分离。这一过程中的错误与肿瘤发生、出生缺陷和细胞死亡有关。在细胞分裂过程中，染色体正确分割的关键是两极纺锤体的建立、排列和分解。这项资助的目的是进一步了解在模式生物酿酒酵母中创造和控制纺锤体组织的蛋白质的结构、组装和动力学。酵母菌的微管组织中心是纺锤体。SPB是一种动态结构，它以细胞周期特有的方式进行重塑。我们已经确定了参与重塑过程的蛋白质，并将对它们进行表征。我们已经提供了SPB的详细建筑描述，并将继续进行结构分析。SPB成核并组织成微管。我们将进行一系列实验，以区分两种当前的成核模型。最后，我们发现动控元件可以在不附着在微管的正端的情况下进行生物定向。在不涉及正端连接的情况下，通过确定动点、马达和其他纺锤体蛋白在双极纺锤体形成中的作用，将获得对双向定向的新见解。由于许多蛋白质和纺锤体特征从酵母到人类都是保守的，我们的研究将向模型提供真核细胞如何正确分配其遗传物质的信息。

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. Accurate chromosome segregation is required for propagation of all cells. Errors in this process are implicated in oncogenesis, birth defects and cell death. Crucial to proper partitioning of the chromosomes during cell division is the establishment, arrangement and then breakdown of a bipolar spindle. The aim of this grant is to further understand the structure, assembly and dynamics of the proteins that create and control the organization of the spindle in the model organism Saccharomyces cerevisiae. The microtubule organizing center in yeast is the spindle pole body (SPB). The SPB is a dynamic structure that undergoes remodeling in a cell-cycle specific manner. We have identified proteins involved in the remodeling process and will characterize them. We have provided a detailed architectural description of the SPB and will continue our structural analysis. The SPB nucleates and organizes microtubules. We will perform a set of experiments designed to distinguish between two current models for nucleation. Finally, we have found that kinetochores can biorient without being attached to the plus-end of microtubules. New insights into biorientation will be obtained from determining the role of kinetochores, motors and other spindle proteins in the formation of the bipolar spindle when plus-end attachment is not involved. Because many proteins and spindle features are conserved from yeast to human, our studies will inform models of how eukaryotic cells correctly distribute their genetic material.