serval:BIB_1366
Elf-1 contributes to the function of the complex interleukin (IL)-2-responsive enhancer in the mouse IL-2 receptor alpha gene.
10.1084/jem.185.7.1211
A1997WU18300007
9104808
Serdobova
I.
author
Pla
M.
author
Reichenbach
P.
author
Sperisen
P.
author
Ghysdael
J.
author
Wilson
A.
author
Freeman
J.
author
Nabholz
M.
author
article
1997
Journal of Experimental Medicine
0022-1007
0022-1007
journal
185
7
1211-1221
Lymphocytes regulate their responsiveness to IL-2 through the transcriptional control of the IL-2R alpha gene, which encodes a component of the high affinity IL-2 receptor. In the mouse IL-2R alpha gene this control is exerted via two regulatable elements, a promoter proximal region, and an IL-2-responsive enhancer (IL-2rE) 1.3 kb upstream. In vitro and in vivo functional analysis of the IL-2rE in the rodent thymic lymphoma-derived, CD4- CD8- cell line PC60 demonstrated that three separate elements, sites I, II, and III, were necessary for IL-2 responsiveness; these three sites demonstrate functional cooperation. Site III contains a consensus binding motif for members of the Ets family of transcription factors. Here we demonstrate that Elf-1, an Ets-like protein, binds to site III and participates in IL-2 responsiveness. In vitro site III forms a complex with a protein constitutively present in nuclear extracts from PC60 cells as well as from normal CD4- CD8- thymocytes. We have identified this molecule as Elf-1 according to a number of criteria. The complex possesses an identical electrophoretic mobility to that formed by recombinant Elf-1 protein and is super-shifted by anti-Elf-1 antibodies. Biotinylated IL-2rE probes precipitate Elf-1 from PC60 extracts provided site III is intact and both recombinant and PC60-derived proteins bind with the same relative affinities to different mutants of site III. In addition, by introducing mutations into the core of the site III Ets-like motif and comparing the corresponding effects on the in vitro binding of Elf-1 and the in vivo IL-2rE activity, we provide strong evidence that Elf-1 is directly involved in IL-2 responsiveness. The nature of the functional cooperativity observed between Elf-1 and the factors binding sites I and II remains unresolved; experiments presented here however suggest that this effect may not require direct interactions between the proteins binding these three elements.
Animals
Base Sequence
Binding Sites
DNA-Binding Proteins/genetics
DNA-Binding Proteins/metabolism
Enhancer Elements, Genetic
Ephrin-A2
Interleukin-2/pharmacology
Mice
Molecular Sequence Data
Mutation
Nuclear Proteins/metabolism
Protein Binding
Proto-Oncogene Proteins/genetics
Proto-Oncogene Proteins c-ets
Receptors, Interleukin-2/genetics
T-Lymphocytes/metabolism
Transcription Factors/genetics
Transcription Factors/metabolism
Tumor Cells, Cultured
eng
60_published
true
University of Lausanne
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