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

Ward, Susan B. and Hernandez-Hoyos, Gabriela and Chen, Fei and Waterman, Marian and Reeves, Raymond and Rothenberg, Ellen V. (1998) Chromatin remodeling of the interleukin-2 gene: distinct alterations in the proximal versus distal enhancer regions. Nucleic Acids Research, 26 (12). pp. 2923-2934. ISSN 0305-1048. PMCID PMC147656.

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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 54 and 34 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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Rothenberg, Ellen V.0000-0002-3901-347X
Additional Information:© 2004 Oxford University Press. Received March 11, 1998; Accepted April 4, 1998. We would like to thank Dan Chen, Paul Garrity, Barbara Wold and Rochelle Diamond for their unwavering advice, interest and support. We are indebted to Eric Davidson, Kai Zinn and Paul Mueller for invaluable comments on the manuscript. We would also like to thank Stephanie Canada for expert technical assistance with the manuscript as well as Jim Staub and Richard Gomez for their help with the figures. Oligonucleotides were provided by the Caltech Biopolymer Synthesis Facility. This work was supported by USPHS grants AI34041 and AG13108 to E.V.R., by the Stowers Institute for Medical Research and, in part, by the State of California Tobacco Related Disease Research Program 4RT-0264.
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Stowers Institute for Medical ResearchUNSPECIFIED
California Tobacco-Related Disease Research Program4RT-0264
Issue or Number:12
PubMed Central ID:PMC147656
Record Number:CaltechAUTHORS:WARnar98
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Usage Policy:No commercial reproduction, distribution, display or performance rights in this work are provided.
ID Code:581
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Deposited On:29 Aug 2005
Last Modified:02 Oct 2019 22:34

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