Novel Egr/NF-AT composite sites mediate activation of the CD95 (APO-1/Fas) ligand promoter in response to T cell stimulation

Min Li-Weber, Oskar Laur, Peter H. Krammer

Research output: Contribution to journalArticle

63 Citations (Scopus)

Abstract

Expression of the CD95 (APO-1/Fas) ligand (CD95L) in activated T cells is a major cause of activation-induced T cell apoptosis. The transcription factors NF-AT and Egr-3 (a member of the immediate-early transcription factors involved in cellular growth and differentiation) have been implicated in activation of the CD95L promoter upon T cell activation. On the basis of DNase I footprinting, electrophoretic mobility shift assay, antibody supershift analysis and transfection studies, we have identified two novel Egr-binding sites 5' upstream of the previously identified Egr site. Mutation analysis of each Egr site shows that all three sites are important for full CD95L promoter activity. Strikingly, all Egr sites, including the previously identified Egr site, are adjacent to or overlap with DNA sequences homologous to NF-AT binding sites and confer T cell activation-induced, cyclosporin A-sensitive transcriptional activity. Antibody supershift analysis revealed that NF-AT and Egr proteins are the components of inducible DNA-binding complexes formed on the two novel Egr sites. Cotransfection experiments showed that Egr-1, Egr-3 and NF-AT display a cooperative and synergistic activation of transcription mediated by these three Egr/NF-AT composite regulatory elements. These findings provide further insight into the mechanisms involved in the regulation of the CD95L expression in response to T cell activation.

Original languageEnglish (US)
Pages (from-to)3017-3027
Number of pages11
JournalEuropean Journal of Immunology
Volume29
Issue number9
DOIs
StatePublished - Jan 1 1999

Fingerprint

Fas Ligand Protein
T-Lymphocytes
NFATC Transcription Factors
Binding Sites
Antibodies
Deoxyribonuclease I
Electrophoretic Mobility Shift Assay
Transcriptional Activation
Cyclosporine
Transfection
Transcription Factors
Apoptosis
Mutation
DNA
Growth

Keywords

  • Apoptosis
  • CD95(APO-1/Fas)L promoter

ASJC Scopus subject areas

  • Immunology and Allergy
  • Immunology

Cite this

Novel Egr/NF-AT composite sites mediate activation of the CD95 (APO-1/Fas) ligand promoter in response to T cell stimulation. / Li-Weber, Min; Laur, Oskar; Krammer, Peter H.

In: European Journal of Immunology, Vol. 29, No. 9, 01.01.1999, p. 3017-3027.

Research output: Contribution to journalArticle

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