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Molecular and Cellular Biology, November 1999, p. 7600-7609, Vol. 19, No. 11
0270-7306/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.

Structural and Functional Cross-Talk between a Distant Enhancer and the epsilon -Globin Gene Promoter Shows Interdependence of the Two Elements in Chromatin

Jennifer C. McDowelldagger and Ann Dean*

Laboratory of Cellular and Developmental Biology, National Institute of Diabetes and Digestive and Kidney Diseases, National Institutes of Health, Bethesda, Maryland 20892-2715

Received 1 June 1999/Returned for modification 21 July 1999/Accepted 2 August 1999

We investigated the requirements for enhancer-promoter communication by using the human beta -globin locus control region (LCR) DNase I-hypersensitive site 2 (HS2) enhancer and the varepsilon -globin gene in chromatinized minichromosomes in erythroid cells. Activation of globin genes during development is accompanied by localized alterations of chromatin structure, and CACCC binding factors and GATA-1, which interact with both globin promoters and the LCR, are believed to be critical for globin gene transcription activation. We found that an HS2 element mutated in its GATA motif failed to remodel the varepsilon -globin promoter or activate transcription yet HS2 nuclease accessibility did not change. Accessibility and transcription were reduced at promoters with mutated GATA-1 or CACCC sites. Strikingly, these mutations also resulted in reduced accessibility at HS2. In the absence of a globin gene, HS2 is similarly resistant to nuclease digestion. In contrast to observations in Saccharomyces cerevisiae, HS2-dependent promoter remodeling was diminished when we mutated the TATA box, crippling transcription. This mutation also reduced HS2 accessibility. The results indicate that the varepsilon -globin promoter and HS2 interact both structurally and functionally and that both upstream activators and the basal transcription apparatus contribute to the interaction. Further, at least in this instance, transcription activation and promoter remodeling by a distant enhancer are not separable.


* Corresponding author. Mailing address: Building 6, Room B1-08, 6 Center Drive, MSC 2715, Bethesda, MD 20892. Phone: (301) 496-6068. Fax: (301) 496-5239. E-mail: anndean{at}helix.nih.gov.

dagger Present address: National Center for Biotechnology Information, National Library of Medicine, National Institutes of Health, Bethesda, MD 20894.


Molecular and Cellular Biology, November 1999, p. 7600-7609, Vol. 19, No. 11
0270-7306/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.



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