GINS MOTION REVEALS REPLICATION FORK PROGRESSION IS REMARKABLY UNIFORM

该子项目是利用资源的众多研究子项目之一 由 NIH/NCRR 资助的中心拨款提供。子项目的主要支持 并且子项目的主要研究者可能是由其他来源提供的， 包括其他 NIH 来源。 子项目可能列出的总成本 代表子项目使用的中心基础设施的估计数量， NCRR 赠款不直接向子项目或子项目工作人员提供资金。 先前的研究已经得出这样的图景：在芽殖酵母基因组的不同部分，DNA 的复制以不同的速率进行。这些先前的实验侧重于新生 DNA 的产生，被解释为暗示复制叉进展的动力学受到局部染色质结构/架构以及与控制转录、修复和表观遗传维持的机制的相互作用的强烈影响。在这里，我们采用了一种补充方法，使用全基因组时间分辨染色质免疫沉淀结合 GINS 复合体（复制叉的一个组成部分）的微阵列分析来分析复制动态。令人惊讶的是，我们的数据表明，无论基因组位置如何，这种复合物都以高度一致的速率进展，这表明酵母中的复制叉动力学比之前设想的更简单、更一致。此外，我们还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分叉运动，并且还揭示了一种微妙的现象，我们将其解释为由低频分叉停滞引起的。 描述这些结果的手稿已发表： M.D. Sekedat、D. Feny¿、R.S. Rogers, A.J.塔克特，J.D. 艾奇森，B.T. Chait“GINS 运动揭示复制叉进展在整个酵母基因组中非常均匀”Mol Systems Biol，6 (2010) 353

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. Primary support for the subproject and the subproject's principal investigator may have been provided by other sources, including other NIH sources. The Total Cost listed for the subproject likely represents the estimated amount of Center infrastructure utilized by the subproject, not direct funding provided by the NCRR grant to the subproject or subproject staff. 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. A manuscript describing these results has been published: M.D. Sekedat, D. Feny¿, R.S. Rogers, A.J. Tackett, J.D. Aitchison, B.T. Chait "GINS Motion Reveals Replication Fork Progression is Remarkably Uniform Throughout the Yeast Genome" Mol Systems Biol, 6 (2010) 353