title: Early growth response proteins (EGR) and nuclear factors of activated T cells (NFAT) form heterodimers and regulate proinflammatory cytokine gene expression.
abstract: Activation of transcription factors by receptor mediated signaling is an essential step for T lymphocyte effector function. Following antigenic stimulation of T cells the two central cytokines IL-2 and TNFalpha are co-expressed and co-regulated. Two important transcription factors, i.e., early growth response (EGR) protein EGR-1 and nuclear factors of activated T cells (NFAT) protein NFATc, regulate transcription of the human IL-2 cytokine and the same combination of EGR and NFAT proteins seems relevant for coordinated cytokine expression. Here we demonstrate that the zinc finger protein EGR-1 and two members of the NFAT protein family bind simultaneously to adjacent elements position -168 to -150 within the TNFalpha promoter. Both promoter sites are important for TNFalpha gene transcription as shown by transfection assays having the IL-2 and TNFalpha promoters linked to a luciferase reporter. The use of promoter deletion constructs with the zinc finger protein (ZIP), the NFAT binding element or a combination of both deleted show a functional cooperation of these elements and of their binding factors. These experiments demonstrate that EGR-1 as well as EGR-4 functionally cooperate with NFAT proteins and induce expression of both cytokine genes. Using tagged NFATc and NFATp in glutathione S-transferase pull down assays showed interaction and physical complex formation of each NFAT protein with recombinant, as well as native, EGR-1 and EGR-4 proteins. Thus EGR-NFAT interaction and complex formation seems essential for human cytokine expression as adjacent ZIP and NFAT elements are conserved in the IL-2 and TNFalpha gene promoters. Binding of regulatory EGR and NFAT factors to these sites and the functional interaction and formation of stable heterodimeric complexes indicate an important role of these factors for gene transcription.
authors.affiliation: Research Group of Biomolecular Medicine, Bernhard Nocht Institute for Tropical Medicine, 20359 Hamburg, Bernhard-Nocht Strasse 74, Germany.
authors.forename: Eva L
authors.initials: EL
authors.lastname: Decker
authors.forename: Nina
authors.initials: N
authors.lastname: Nehmann
authors.forename: Eva
authors.initials: E
authors.lastname: Kampen
authors.forename: Hermann
authors.initials: H
authors.lastname: Eibel
authors.forename: Peter F
authors.initials: PF
authors.lastname: Zipfel
authors.forename: Christine
authors.initials: C
authors.lastname: Skerka
mesh_terms: D001483:Base Sequence; D001665:Binding Sites; D002460:Cell Line; D017124:Conserved Sequence; D016207:Cytokines / Q000235:genetics*; D004268:DNA-Binding Proteins / Q000378:metabolism / Q000502:physiology*; D019281:Dimerization; D051766:Early Growth Response Protein 1; D051765:Early Growth Response Transcription Factors; D006801:Humans; D017874:Immediate-Early Proteins; D007249:Inflammation / Q000235:genetics / Q000276:immunology; D007376:Interleukin-2 / Q000235:genetics; D019169:Jurkat Cells; D050778:NFATC Transcription Factors; D009687:Nuclear Proteins; D011401:Promoter Regions, Genetic; D020218:Response Elements; D014157:Transcription Factors / Q000378:metabolism / Q000502:physiology*; D014158:Transcription, Genetic; D015533:Transcriptional Activation; D014409:Tumor Necrosis Factor-alpha / Q000235:genetics
chemical_list: D016207:Cytokines; D004268:DNA-Binding Proteins; C497325:EGR1 protein, human; C111028:EGR4 protein, human; D051766:Early Growth Response Protein 1; D051765:Early Growth Response Transcription Factors; D017874:Immediate-Early Proteins; D007376:Interleukin-2; D050778:NFATC Transcription Factors; D009687:Nuclear Proteins; D014157:Transcription Factors; D014409:Tumor Necrosis Factor-alpha