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J Biol Chem, Vol. 274, Issue 49, 35203-35210, December 3, 1999

Transcriptional Regulation of Fas Gene Expression by GA-binding Protein and AP-1 in T Cell Antigen Receptor·CD3 Complex-stimulated T Cells

Xiao Rui LiDagger , Anita S.-F. ChongDagger , Jianming Wuparallel , Kenneth A. Roebuck, Aseem Kumar**, Joseph E. Parrillo**, Ulf R. RappDagger Dagger , Robert P. Kimberlyparallel , James W. WilliamsDagger , and Xiulong XuDagger

From the Departments of Dagger  General Surgery,  Immunology/Microbiology, and ** Medicine, Rush Presbyterian St. Luke's Medical Center, Chicago, Illinois 60612, the parallel  Division of Clinical Immunology and Rheumatology, Department of Medicine, School of Medicine, University of Alabama at Birmingham, Birmingham, Alabama 35284, and the Dagger Dagger  Institute of Medical Radiation Research and Cell Biology, University of Würzburg, D-97080 Würzburg, Germany

Fas (CD95 or APO-1), a transmembrane cell surface receptor of the tumor necrosis factor receptor family, is up-regulated in activated T lymphocytes. Our present study identified an upstream enhancer element (between nucleotide positions -862 and -682) containing a GA-binding protein (GABP) site and a low affinity activating protein-1 (AP-1)-binding site. T cell activation increased the DNA binding of GABP and AP-1 to this enhancer site. The specificity of GABP and AP-1 binding was demonstrated by competition electrophoretic mobility shift assay and supershift electrophoretic mobility shift assay with antibodies against GABP and AP-1, respectively. Mutational analysis of Fas promoter revealed that both GABP- and AP-1-binding sites were required for initiating Fas gene transcription. We further show that anti-CD3 mAb, phorbol 12-myristate 13-acetate, and phorbol 12-myristate 13-acetate/ionomycin strongly activated promoters carrying multiple copies of the Fas enhancer, and mutation of either the GABP or AP-1 binding site severely reduced transcriptional activity. Taken together, these results suggest that the transcription factors GABP and AP-1 play a critical role in the induction of Fas gene expression in T cell antigen receptor·CD3-stimulated Jurkat cells.

Copyright © 1999 by The American Society for Biochemistry and Molecular Biology, Inc.
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