Laboratory of Molecular and Cellular Biology, Graduate School of Bioresources, Mie University, Tsu, Japan.
Tsuji Health and Beauty Science Laboratory, Mie University, Tsu, Japan.
Cytogenet Genome Res. 2021;161(8-9):437-444. doi: 10.1159/000518263. Epub 2021 Sep 28.
E/L Repli-seq is a powerful tool for detecting cell type-specific replication landscapes in mammalian cells, but its potential to monitor DNA replication under replication stress awaits better understanding. Here, we used E/L Repli-seq to examine the temporal order of DNA replication in human retinal pigment epithelium cells treated with the topoisomerase I inhibitor camptothecin. We found that the replication profiles by E/L Repli-seq exhibit characteristic patterns after replication-stress induction, including the loss of specific initiation zones within individual early replication timing domains. We also observed global disappearance of the replication timing domain structures in the profiles, which can be explained by checkpoint-dependent suppression of replication initiation. Thus, our results demonstrate the effectiveness of E/L Repli-seq at identifying cells with replication-stress-induced altered DNA replication programs.
E/L Repli-seq 是一种强大的工具，可用于检测哺乳动物细胞中特定细胞类型的复制景观，但它在复制应激下监测 DNA 复制的潜力仍有待更好地理解。在这里，我们使用 E/L Repli-seq 来研究用拓扑异构酶 I 抑制剂喜树碱处理的人视网膜色素上皮细胞中的 DNA 复制的时间顺序。我们发现，E/L Repli-seq 的复制图谱在复制应激诱导后表现出特征性模式，包括单个早期复制起始时间域内特定起始区的丢失。我们还观察到图谱中复制起始时间域结构的全局消失，这可以通过复制起始的检查点依赖性抑制来解释。因此，我们的结果表明 E/L Repli-seq 能够有效地识别具有复制应激诱导的改变的 DNA 复制程序的细胞。
