Epner E, Forrester W C, Groudine M
Fred Hutchinson Cancer Research Center, Seattle, WA 98104.
Proc Natl Acad Sci U S A. 1988 Nov;85(21):8081-5. doi: 10.1073/pnas.85.21.8081.
The timing of DNA replication of the human beta-globin gene locus has been studied by blot hybridization of newly synthesized BrdUrd-substituted DNA from cells in different stages of the S phase. Using probes that span greater than 120 kilobases across the human beta-globin gene locus, we show that the majority of this domain replicates in early S phase in the human erythroleukemia cell line K562 and in middle-to-late S phase in the lymphoid cell line Manca. However, in K562 cells three small regions display a strikingly different replication pattern than adjacent sequences. These islands, located in the inter-gamma-globin gene region and approximately 20 kilobases 5' to the epsilon-globin gene and 20 kilobases 3' to the beta-globin gene, replicate later and throughout S phase. A similar area is also present in the alpha-globin gene region in K562 cells. We suggest that these regions may represent sites of termination of replication forks.
通过对处于S期不同阶段的细胞中新合成的BrdUrd取代DNA进行印迹杂交,研究了人类β-珠蛋白基因座DNA复制的时间。使用跨越人类β-珠蛋白基因座超过120千碱基的探针,我们发现该结构域的大部分在人类红白血病细胞系K562的S期早期复制,而在淋巴样细胞系Manca中则在S期中后期复制。然而,在K562细胞中,三个小区域显示出与相邻序列明显不同的复制模式。这些区域位于γ-珠蛋白基因间区域以及ε-珠蛋白基因5'端约20千碱基和β-珠蛋白基因3'端约20千碱基处,在整个S期后期复制。K562细胞的α-珠蛋白基因区域也存在类似区域。我们认为这些区域可能代表复制叉的终止位点。