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J. Biol. Chem., Vol. 279, Issue 48, 50350-50357, November 26, 2004

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Recruitment of Transcription Complexes to the -Globin Gene Locus in Vivo and in Vitro^*

Karen F. Vieira, Padraic P. Levings, Meredith A. Hill, Valerie J. Crusselle, Sung-Hae Lee Kang, James Douglas Engel¶, and Jörg Bungert||

From the Department of Biochemistry and Molecular Biology, Centers for Gene Therapy and Mammalian Genetics, College of Medicine, University of Florida, Gainesville, Florida 32610 and ^¶Department of Cell and Developmental Biology, University of Michigan, Ann Arbor, Michigan 48109-0616

Erythroid-specific, high level expression of the -globin genes is regulated by the locus control region (LCR), composed of multiple DNase I-hypersensitive sites and located far upstream of the genes. Recent studies have shown that LCR core elements recruit RNA polymerase II (pol II). In the present study we demonstrate the following: 1) pol II and other basal transcription factors are recruited to LCR core hypersensitive elements; 2) pol II dissociates from and re-associates with the globin gene locus during replication; 3) pol II interacts with the LCR but not with the -globin gene prior to erythroid differentiation in embryonic stem cells; and 4) the erythroid transcription factor NF-E2 facilitates the transfer of pol II from immobilized LCR constructs to a -globin gene in vitro. The data are consistent with the hypothesis that the LCR serves as the primary attachment site for the recruitment of macromolecular complexes involved in chromatin structure alterations and transcription of the globin genes.

Received for publication, August 4, 2004 , and in revised form, September 2, 2004.

^* This work was supported by National Institutes of Health Grants DK058209 and DK0522356 (to J. B.). The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

Both authors contributed equally to this work.

^|| To whom correspondence should be addressed: Dept. of Biochemistry and Molecular Biology, University of Florida College of Medicine, P. O. Box 100245, 1600 SW Archer Rd., Gainesville, FL 32610. Tel.: 352-392-0121; Fax: 352-392-2953; E-mail: jbungert{at}college.med.ufl.edu.

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