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Nucleic Acids Research, Vol 26, Issue 12 2923-2934, Copyright © 1998 by Oxford University Press

ARTICLES

Chromatin remodeling of the interleukin-2 gene: distinct alterations in the proximal versus distal enhancer regions

SB Ward, G Hernandez-Hoyos, F Chen, M Waterman, R Reeves and EV Rothenberg
Division of Biology MC156-29, California Institute of Technology, Pasadena, CA 91125, USA.

Known transcription factor-DNA interactions in the minimal enhancer of the murine interleukin-2 gene (IL-2) do not easily explain the T cell specificity of IL-2 regulation. To seek additional determinants of cell type specificity, in vivo methodologies were employed to examine chromatin structure 5' and 3' of the 300 bp IL-2 proximal promoter/enhancer region. Restriction enzyme accessibility revealed that until stimulation the IL-2 proximal promoter/enhancer exists in a closed conformation in resting T and non-T cells alike. Within this promoter region, DMS and DNase I genomic footprinting also showed no tissue-specific differences prior to stimulation. However, DNase I footprinting of the distal -600 to -300 bp region revealed multiple tissue-specific and stimulation-independent DNase I hypersensitive sites. Gel shift assays detected T cell-specific complexes binding within this region, which include TCF/LEF or HMG family and probable Oct family components. Upon stimulation, new DNase I hypersensitive sites appeared in both the proximal and distal enhancer regions, implying that there may be a functional interaction between these two domains. These studies indicate that a region outside the established IL-2 minimal enhancer may serve as a stable nucleation site for tissue- specific factors and as a potential initiation site for activation- dependent chromatin remodeling.
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