STRUCTURAL ANALYSIS OF A EUKARYOTIC CENTEROMERE

真核染色体着丝粒是特定的区域 沿着染色质纤维，它在 细胞分裂过程中的染色体运动。着丝粒DNA 从酵母中分离的序列，酿酒酵母， 使导入酵母的外源DNA发挥普通作用 酵母细胞分裂过程中的染色体。我们将研究如何 着丝粒DNA序列与染色质成分相互作用 在细胞核中，包括核蛋白和微管，以 产生了一个有功能的动粒。我们将确定何时 动粒是在细胞周期中组装的，以及这是如何 结构在细胞分裂期间保持不变。我们将研究 动粒的生物化学和分子组成 使用分子技术将该复合体从 染色质纤维。与此相关的蛋白质 体内的结构将被确定。我们会提高抗体 以确定它们在细胞内的定位， 以及分离编码DNA序列的蛋白质。我们会 分离着丝粒突变体作为一种替代方法 鉴定正常染色体所需的基因产物 种族隔离。识别所需的反式作用因子 因为染色体运动将使我们能够确定这些 这些因子与着丝粒DNA相互作用，从而产生 参与染色体运动的功能单位。知识 控制着丝粒组织的因素 可能提供了管理分子的基本原理 细胞分裂的机制。 细胞分裂是生物体的中心过程。中的更改 细胞分裂的控制可能是一个重要的机制 导致异常细胞群体的产生，包括 癌细胞。着丝粒异常，早熟着丝粒 在以下情况下发现了分离和染色体不稳定 Roberts‘s-SC青光眼、共济失调毛细血管扩张症和一些 其他综合征。对细胞控制因素的认识 除法可能是进一步理解 这些功能障碍。

Centromeres of eukaryotic chromosomes are specific regions along the chromatin fiber that play a fundamental role in chromosome movement during cell division. Centromere DNA sequences isolated from the yeast, Saccharomyces cerevisiae, enable foreign DNA introduced into yeast to function as ordinary yeast chromosomes during cell division. We will examine how the centromere DNA sequence interacts with chromatin components in the cell nucleus, including nuclear proteins and microtubules, to give rise to a functional kinetochore. We will determine when the kinetochore is assembled during the cell cycle, and how this structure is maintained during cell division . We will examine the biochemical and molecular constitution of the kinetochore by using molecular techniques to detach this complex from the chromatin fiber. The proteins that are associated with this structure in vivo will the be identified. We will raise antibodies against the proteins to determine their intracellular localization, as well as to isolate the protein coding DNA sequences. We will isolate mutants in centromere function as an alternative method to identify gene products required for proper chromosome segregation. The identification of trans-acting factors required for chromosome movement will enable us to determine how these factors interact with the centromere DNA to give rise to a functional unit involved in chromosome locomotion. Knowledge of the factors that control the organization of the centromere may provide fundamental principles governing the molecular mechanism of cell division. Cell division is a central process of living organisms. Changes in the control of cell division may be an important mechanism that results in the production of aberrant cell populations, including cancer cells. Centromeric abnormalities, premature centromere separation and chromosome instability have been found in cases of Roberts' - SC phocomelia, ataxia telangiectasia, and a number of other syndromes. An understanding of the factors controlling cell division may be fundamental for the further understanding of these dysfunctions.