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Molecular and Cellular Biology, January 2008, p. 575-586, Vol. 28, No. 2
0270-7306/08/$08.00+0     doi:10.1128/MCB.00943-07
Copyright © 2008, American Society for Microbiology. All Rights Reserved.

GATA-1 Modulates the Chromatin Structure and Activity of the Chicken {alpha}-Globin 3' Enhancer{triangledown} ,{dagger}

Martín Escamilla-Del-Arenal and Félix Recillas-Targa*

Instituto de Fisiología Celular, Departamento de Genética Molecular, Universidad Nacional Autónoma de México, Apartado Postal 70-242, México D.F. 04510, México

Received 28 May 2007/ Returned for modification 31 August 2007/ Accepted 23 October 2007

Long-distance regulatory elements and local chromatin structure are critical for proper regulation of gene expression. Here we characterize the chromatin conformation of the chicken {alpha}-globin silencer-enhancer elements located 3' of the domain. We found a characteristic and erythrocyte-specific structure between the previously defined silencer and the enhancer, defined by two nuclease hypersensitive sites, which appear when the enhancer is active during erythroid differentiation. Fine mapping of these sites demonstrates the absence of a positioned nucleosome and the association of GATA-1. Functional analyses of episomal vectors, as well as stably integrated constructs, revealed that GATA-1 plays a major role in defining both the chromatin structure and the enhancer activity. We detected a progressive enrichment of histone acetylation on critical enhancer nuclear factor binding sites, in correlation with the formation of an apparent nucleosome-free region. On the basis of these results, we propose that the local chromatin structure of the chicken {alpha}-globin enhancer plays a central role in its capacity to differentially regulate {alpha}-globin gene expression during erythroid differentiation and development.

* Corresponding author. Mailing address: Instituto de Fisiología Celular, Departamento de Genética Molecular, Universidad Nacional Autónoma de México, Apartado Postal 70-242, México D.F. 04510, México. Phone: (52 55) 56 22 56 74. Fax: (52 55) 56 22 56 30. E-mail: frecilla{at}ifc.unam.mx

{triangledown} Published ahead of print on 5 November 2007.

{dagger} Supplemental material for this article may be found at http://mcb.asm.org/.

Molecular and Cellular Biology, January 2008, p. 575-586, Vol. 28, No. 2
0270-7306/08/$08.00+0     doi:10.1128/MCB.00943-07
Copyright © 2008, American Society for Microbiology. All Rights Reserved.



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