NFAT but not NF-kappaB is critical for transcriptional induction of the prosurvival gene A1 after IgE receptor activation in mast cells.
FcepsilonRI-activation-induced survival of mast cells is dependent on the expression and function of the prosurvival protein A1. The expression of A1 in lymphocytes and monocytes has previously been described to be transcriptionally regulated by NF-kappaB. Here we demonstrate that the expression of A1 in mast cells is not dependent on NF-kappaB but that NFAT plays a crucial role. FcepsilonRI-induced A1 expression was not affected in mast cells overexpressing an IkappaB-alpha super-repressor or cells lacking NF-kappaB subunits RelA, c-Rel, or c-Rel plus NF-kappaB1 p50. In contrast, inhibition of calcineurin and NFAT by cyclosporin A abrogated the expression of A1 in mast cells on FcepsilonRI-activation but had no effect on lipopolysaccharide-induced expression of A1 in J774A.1 monocytic cells. Cyclosporin A also inhibited luciferase expression in an A1 promoter reporter assay. A putative NFAT binding site in the A1 promoter showed inducible protein binding after FcepsilonRI crosslinking or treatment with ionomycin as detected in a band shift assay or chromatin immunoprecipitation. The binding protein was identified as NFAT1. Finally, mast cells expressing constitutively active NFAT1 exhibit increased expression of A1 after FcepsilonRI-stimulation. These results indicate that, in FcepsilonRI stimulated mast cells, A1 is transcriptionally regulated by NFAT1 but not by NF-kappaB.
Ullerås
Erik
E
Clinical Immunology and Allergy Unit, Department of Medicine, Karolinska Institutet, Stockholm, Sweden.
Karlberg
Mats
M
Möller Westerberg
Christine
C
Alfredsson
Jessica
J
Gerondakis
Steve
S
Strasser
Andreas
A
Nilsson
Gunnar
G
D000818:Animals; D002470:Cell Survival; D002478:Cells, Cultured; D003429:Cross Reactions / Q000276:immunology; D016572:Cyclosporine / Q000494:pharmacology; D015759:Ionomycin / Q000494:pharmacology; D008264:Macrophages / Q000187:drug effects; D008407:Mast Cells / Q000166:cytology / Q000187:drug effects / Q000276:immunology* / Q000378:metabolism*; D051379:Mice; D008810:Mice, Inbred C57BL; D018345:Mice, Knockout; D015778:Minor Histocompatibility Antigens; D016328:NF-kappa B / Q000172:deficiency / Q000235:genetics / Q000378:metabolism; D050778:NFATC Transcription Factors / Q000235:genetics / Q000378:metabolism*; D011401:Promoter Regions, Genetic / Q000235:genetics; D019253:Proto-Oncogene Proteins c-bcl-2 / Q000235:genetics* / Q000378:metabolism*; D017455:Receptors, IgE / Q000276:immunology*; D014158:Transcription, Genetic / Q000235:genetics*
C096816:BCL2-related protein A1; D015778:Minor Histocompatibility Antigens; D016328:NF-kappa B; D050778:NFATC Transcription Factors; D019253:Proto-Oncogene Proteins c-bcl-2; D017455:Receptors, IgE; D015759:Ionomycin; D016572:Cyclosporine