Laboratory for Gaseous Ion Chemistry, The Rockefeller University, New York, NY 10065, USA.
Mol Syst Biol. 2010;6:353. doi: 10.1038/msb.2010.8. Epub 2010 Mar 9.
Previous studies have led to a picture wherein the replication of DNA progresses at variable rates over different parts of the budding yeast genome. These prior experiments, focused on production of nascent DNA, have been interpreted to imply that the dynamics of replication fork progression are strongly affected by local chromatin structure/architecture, and by interaction with machineries controlling transcription, repair and epigenetic maintenance. Here, we adopted a complementary approach for assaying replication dynamics using whole genome time-resolved chromatin immunoprecipitation combined with microarray analysis of the GINS complex, an integral member of the replication fork. Surprisingly, our data show that this complex progresses at highly uniform rates regardless of genomic location, revealing that replication fork dynamics in yeast is simpler and more uniform than previously envisaged. In addition, we show how the synergistic use of experiment and modeling leads to novel biological insights. In particular, a parsimonious model allowed us to accurately simulate fork movement throughout the genome and also revealed a subtle phenomenon, which we interpret as arising from low-frequency fork arrest.
在出芽酵母基因组的不同部位,DNA 的复制以不同的速度进行。这些之前的实验集中在新生 DNA 的产生上,被解释为意味着复制叉推进的动力学受到局部染色质结构/架构的强烈影响,并受到控制转录、修复和表观遗传维持的机制的影响。在这里,我们采用了一种互补的方法来检测使用全基因组时间分辨染色质免疫沉淀结合微阵列分析 GINS 复合物(复制叉的一个组成部分)的复制动力学。令人惊讶的是,我们的数据表明,无论基因组位置如何,该复合物都以高度均匀的速度前进,这表明酵母中的复制叉动力学比以前想象的更简单、更均匀。此外,我们展示了实验和建模的协同使用如何带来新的生物学见解。特别是,一个简约的模型使我们能够准确地模拟整个基因组中的叉运动,也揭示了一个微妙的现象,我们将其解释为源于低频叉暂停。