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Properties of transcription factors regulating interleukin-2 gene
transcription through the NFAT binding site in untreated or drug- treated
naive and memory T-helper cells
A Mouzaki and D Rungger
Department of Medicine, Geneva University Hospital, Switzerland.
Combining in vitro DNA binding studies and functional transcription assays
in the Xenopus oocyte, we have tested the presence and functional state of
transcription factors controlling the interleukin-2 (IL-2) promoter through
the NFAT binding site. In naive T-helper cells, the IL-2 gene is repressed
by a silencer. After first mitogenic stimulation, this silencer becomes
undetectable while an activator is newly synthesized. In resting memory
cells, the activator has low DNA- binding affinity and is located in the
cytoplasm. However, no silencer is formed. Upon renewed cellular
activation, this pre-existing activator is again targeted to the nucleus
and regains function in promoting transcription. Cyclosporin A and FK506
act on two distinct levels of the IL-2 control mechanism. They prevent
nuclear transport and reactivation of the performed activator in memory
cells and, in naive cells, they render the silencer resistant to
displacement by the activator. DNA-binding of silencer and activator from
T-helper, and NFAT-1 from Jurkat cells, requires the same three G residues,
but cross- linking analyses show differences in their constituent subunits.
Supershift experiments show that the activator contains fra-2 and junD,
whereas the silencer reacts with none of the antibodies tested.
Volume 84,
Issue 8,
pp. 2612-2621,
10/15/1994
Copyright © 1994 by The American Society of Hematology
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