Abstract:
DNA replication ensures the complete and accurate duplication of the genome. The traditional approach to analysing perturbation of DNA replication is to use chemical inhibitors, such as hydroxyurea or aphidicolin, that slow or stall replication fork progression throughout the genome. An alternative approach is to perturb replication at a single site in the genome that permits a more forensic investigation of the cellular response to the stalling or disruption of a replication fork. This has been achieved in several organisms using different systems that share the common feature of utilizing the high affinity binding of a protein to a defined DNA sequence that is integrated into a specific locus in the host genome. Protein-mediated replication fork blocking systems of this sort have proven very valuable in defining how cells cope with encountering a barrier to fork progression. In this review, we compare protein-based replication fork barrier systems from different organisms that have been developed to generate site-specific replication fork perturbation.
摘要:
DNA复制确保基因组的完整和准确复制。分析DNA复制扰动的传统方法是使用化学抑制剂,如羟基脲或蚜虫霉素,在整个基因组中缓慢或停滞的复制叉进展。另一种方法是在基因组中的单个位点干扰复制,这允许对复制叉的停滞或破坏进行更法医的细胞反应调查。这已经在使用不同系统的几种生物体中实现,所述不同系统具有利用蛋白质与被整合到宿主基因组中的特定基因座中的确定的DNA序列的高亲和力结合的共同特征。这种蛋白质介导的复制叉阻断系统已被证明在定义细胞如何应对叉进展障碍方面非常有价值。在这次审查中,我们比较了来自不同生物的基于蛋白质的复制叉屏障系统,这些生物已被开发用于产生位点特异性复制叉扰动。