Activation and Repression of a β-Globin Gene in Cell Hybrids Is Accompanied by a Shift in Its Temporal Replication

Abstract

To investigate whether a switch in the transcriptional activity of a gene is associated with a change in the timing of replication during the S phase, we examined the replication timing of the β-globin genes in two different types of somatic cell hybrids. In mouse hepatoma (Hepa la) x mouse erythroleukemia (MEL) hybrid cells, the β-globin gene from the MEL parent is transcriptionally inactivated and is later replicating than in the parental MEL cell line. In human fibroblast (GM3552) x MEL hybrid cells, the human β-globin gene is transcriptionally activated, and all of the sequences within the human β-globin domain (200 kilobases) we have examined appear to be earlier replicating than those in the parental fibroblast cell line. The chromatin configuration of the activated human β-globin domain in the hybrids is relatively more sensitive to nucleases than that in the fibroblasts. Furthermore, major nuclease-hypersensitive sites that were absent in the chromatin flanking the distal 5′ region of the human β-globin gene cluster in the parental fibroblast cell line are present in the transcriptionally activated domain in the hybrid cell line. These results suggest that timing of replication of globin genes has been altered in these hybrid cells and thus is not fixed during the process of differentiation.