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Molecular and Cellular Biology, October 1998, p. 5992-6000, Vol. 18, No. 10
0270-7306/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.

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

Andreas Reik,¹ Agnes Telling,¹ Galynn Zitnik,¹ Daniel Cimbora,¹ Elliot Epner,^1, and Mark Groudine^1,2,*

Division of Basic Science, Fred Hutchinson Cancer Research Center, Seattle, Washington 98109,¹ and Department of Radiation Oncology, University of Washington Medical School, Seattle, Washington 98195²

Received 19 May 1998/Accepted 30 June 1998

Studies in many systems have led to the model that the human -globin locus control region (LCR) regulates the transcription, chromatin structure, and replication properties of the -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 -globin locus in an erythroid background, transcription of all -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 -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 -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 -like globin genes.

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^* Corresponding author. Mailing address: Division of Basic Science, Fred Hutchinson Cancer Research Center, 1100 Fairview Ave. N., Seattle, WA 98109. Phone: (206) 667-4497. Fax: (206) 667-5894. E-mail: markg{at}fhcrc.org.

Present address: Department of Medicine, Arizona Cancer Center, University of Arizona, Tucson, AZ 85724.

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Molecular and Cellular Biology, October 1998, p. 5992-6000, Vol. 18, No. 10
0270-7306/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.

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