TARGETING AND FUNCTION OF CENTROMERE PROTEIN A

有丝分裂和减数分裂时的染色体分离对所有人都是必不可少的。 细胞繁殖。这一进程的中断具有深远的影响 对人类健康的影响。着丝粒指定程序集 染色质纤维上的一组独特的功能元件 提供动力、姐妹染色单体凝聚力和机械感觉 将这些功能与细胞周期检查点联系起来的机制。 CENP-A是核心的着丝粒特异性同系物 核小体蛋白质组蛋白H3。CENP-A的特异性组装 人类着丝粒需要蛋白质-蛋白质的结合 核小体内的相互作用，分化的DNA接触点 在核小体的表面和一个独特的调控途径上， 导致着丝粒选择性识别的假说 DNA通过CENP-A组蛋白复合体。CENP-A核小体将 用亲和力分离的着丝粒DNA序列重组 对于CENP-A。对DNA-蛋白质相互作用的分析将揭示 CENP-A是否表现出对着丝粒DNA的特异性识别。 CENP-A在染色质中的组装将被检查以测试 CENP-A染色质组装通过一种 新颖，非S时相通路。CENP-A相关蛋白将是 分离和表征以探索其功能相互作用。 突变方法将结合长期的转基因方法 用新的着丝粒运动性、机械性测定方法 活细胞中的属性和空间组织。

Chromosome segregation at mitosis and meiosis is essential for all cellular reproduction. Disruptions in this process have profound consequences for human health. Centromeres specify the assembly of a unique set of functional elements on the chromatin fiber that provide motility, sister chromatid cohesion and mechanosensory mechanisms that link these functions to cell cycle checkpoints. CENP-A is a centromere specific homologue of the core nucleosomal protein histone H3. The specific assembly of CENP-A at human centromeres requires a combination of protein-protein interactions within the nucleosome, differentiated DNA contact sites at the surface of the nucleosome and a unique regulatory pathway, leading to a hypothesis for selective recognition of centromeric DNA by CENP-A histone complexes. CENP-A nucleosomes will reconstituted with centromeric DNA sequences isolated by affinity for CENP-A. Analysis of DNA-protein interactions will reveal whether CENP-A exhibits specific recognition of centromeric DNA. The assembly of CENP-A into chromatin will be examined to test the hypothesis that CENP-A chromatin assembly occurs through a novel, non-S phase pathway. CENP-A associated proteins will be isolated and characterized to explore its functional interactions. Mutational approaches will combine long term transfection methods with novel assay methods for centromere motility, mechanical properties and spatial organization in living cells.