The Locus Control Region Is Necessary for Gene Expression in the Human β-Globin Locus but Not the Maintenance of an Open Chromatin Structure in Erythroid Cells

ABSTRACT

Studies in many systems have led to the model that the human β-globin locus control region (LCR) regulates the transcription, chromatin structure, and replication properties of the β-globin locus. However the precise mechanisms of this regulation are unknown. We have developed strategies to use homologous recombination in a tissue culture system to examine how the LCR regulates the locus in its natural chromosomal environment. Our results show that when the functional components of the LCR, as defined by transfection and transgenic studies, are deleted from the endogenous β-globin locus in an erythroid background, transcription of all β-globin genes is abolished in every cell. However, formation of the remaining hypersensitive site(s) of the LCR and the presence of a DNase I-sensitive structure of the β-globin locus are not affected by the deletion. In contrast, deletion of 5′HS5 of the LCR, which has been suggested to serve as an insulator, has only a minor effect on β-globin transcription and does not influence the chromatin structure of the locus. These results show that the LCR as currently defined is not necessary to keep the locus in an “open” conformation in erythroid cells and that even in an erythroid environment an open locus is not sufficient to permit transcription of the β-like globin genes.

ACKNOWLEDGMENTS

We thank Michael Bender, Mike Bulger, and Steve Fiering for critical reading of the manuscript, and we thank the Hutchinson Cancer Center Image Analysis Laboratory for assistance in PhosphorImager analysis.

This work was supported by fellowships from the Deutsche Forschungsgemeinschaft and the Leukemia Research Foundation to A.R., a fellowship from the American Cancer Society to D.C., and National Cancer Institute grant CA54337 and NIH grants DK52854 and DK 44746 to M.G.