Divison of Basic Sciences, Fred Hutchinson Cancer Research Center, Seattle, Washington, USA.
Nat Struct Mol Biol. 2010 Apr;17(4):430-7. doi: 10.1038/nsmb.1780. Epub 2010 Mar 14.
Although histone modifications have been implicated in many DNA-dependent processes, their precise role in DNA replication remains largely unknown. Here we describe an efficient single-step method to specifically purify histones located around an origin of replication from Saccharomyces cerevisiae. Using high-resolution MS, we have obtained a comprehensive view of the histone modifications surrounding the origin of replication throughout the cell cycle. We have discovered that acetylation of histone H3 and H4 is dynamically regulated around an origin of replication, at the level of multiply acetylated histones. Furthermore, we find that this acetylation is required for efficient origin activation during S phase.
尽管组蛋白修饰与许多依赖于 DNA 的过程有关，但它们在 DNA 复制中的确切作用在很大程度上仍是未知的。在这里，我们描述了一种从酿酒酵母中特异性纯化复制起始点周围组蛋白的高效单步方法。通过高分辨率 MS，我们在整个细胞周期中获得了围绕复制起始点的组蛋白修饰的全面视图。我们发现，在复制起始点周围，组蛋白 H3 和 H4 的乙酰化水平是动态调节的，多乙酰化组蛋白的水平也发生了变化。此外，我们发现这种乙酰化对于 S 期有效激活起始点是必需的。
