Compositional and structural analysis of DNA replication origins by locus-specific chromatin isolation (A29*)

染色质动力学和局部染色质组织对DNA复制起点的活性有重要贡献，但精确的染色质和结构特征，如确切的核小体构型、组蛋白修饰状态或允许有效和及时复制起始的额外染色质因子的要求，在很大程度上仍然是难以捉摸的。在这个项目中，我们将使用位点特异性染色分离，以针对酵母染色体III上的四个不同的复制起点，这些复制起点在时间和发射效率方面的内在复制特性方面存在重大差异。生物化学和功能分析的组合，提出了研究是否本地染色质结构决定这些地区的能力，启动DNA复制在体内。

Chromatin dynamics and the local chromatin organization critically contribute to the activity of DNA replication origins, but the precise chromatin and structural features like the exact nucleosome configuration, histone modification state or requirement of additional chromatin factors to allow efficient and timely replication initiation remain largely elusive. In this project, we will use locus-specific chromation isolation to target four distinct replication origins on yeast chromosome III that show major differences in their intrinsic replication properties regarding timing and firing efficiency. A combination of biochemical and functional assays is proposed to study whether the local chromatin structure determines the capacity of these regions to initiate DNA replication in vivo.