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J. Biol. Chem., Vol. 275, Issue 47, 36605-36611, November 24, 2000
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From the Expression of cytokine genes in T cells is
thought to result from a complex network of antigen- and
mitogen-activated transcriptional regulators. CP2, a factor homologous
to Drosophila Elf-1 and previously found to be a critical
regulator of several viral and cellular genes in response to
developmental signals, is rapidly activated in T helper (Th) cells in
response to mitogenic stimulation. Here we show that overexpression of
CP2 enhances interleukin (IL)-4 promoter-driven chloramphenicol
acetyltransferase expression, while repressing IL-2 promoter activity,
in transiently transfected Jurkat cells. A CP2-protected element,
partially overlapping the nuclear factor of activated T cell-binding P2
sequence, was required for IL-4 promoter activation in
CP2-overexpressing Jurkat cells. This CP2-response element is
the site of a cooperative interaction between CP2 and an inducible
heteromeric co-factor(s). Mutation of conserved nucleotide contacts
within the CP2-response element prevented CP2 binding and significantly
reduced constitutive and induced IL-4 promoter activity. Expression of
a CP2 mutant lacking the Elf-1-homology region of the DNA-binding
domain inhibited IL-4 promoter activity in a dominant negative fashion
in transiently transfected Jurkat cells. Moreover, overexpressed CP2
markedly enhanced, while its dominant negative mutant consistently
suppressed, expression of the endogenous IL-4 gene in the murine Th2
cell line D10. Taken together, these findings point to CP2 as a
critical IL-4 transactivator in Th cells.
Identification and Characterization of a Critical CP2-binding
Element in the Human Interleukin-4 Promoter*
§,
,,,,, and Department of Medicine, The Johns Hopkins
School of Medicine, Baltimore, Maryland 21224, the ¶ Schepens Eye
Research Institute and Committee on Immunology and Division of
Rheumatology, Immunology, and Allergy, Department of Medicine, Brigham
and Women's Hospital, Harvard Medical School, Boston, Massachusetts
02114, and the Molecular Biology Program and Graduate School of
Medical Sciences, Cornell University, Memorial Sloan-Kettering Cancer
Center, New York, New York 10021
*
This work was supported by National Institutes of Health
Grants AI41463 (to V. C.), AI01152 (to S. N. G.), GM49661, EY1901, EY12523, and CA89559 (to S. J. O.).The costs of publication of this
article were defrayed in part by the
payment of page charges. The article
must therefore be hereby marked
"advertisement" in accordance with 18 U.S.C. Section
1734 solely to indicate this fact.
Copyright © 2000 by The American Society for Biochemistry and Molecular Biology, Inc.
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