Functional synergism of STAT6 with either NF-kappa B or PU.1 to mediate IL-4-induced activation of IgE germline gene transcription.
Ig heavy chain class switching to IgE is directed by IL-4 and IL-13 by inducing transcription from the IgE germline promoter. A crucial transcription factor in this process is STAT6, which binds to a specific DNA element upon cytokine activation. In this paper it is shown that the B cell- and monocyte-specific factor PU.1 interacts with a closely spaced sequence in the human IgE germline promoter that overlaps with a previously described binding site for NF kappa B/rel. The authenticity of PU.1 was demonstrated by specific competition and supershifts in EMSA experiments. In addition, in vitro translated PU.1 could interact with an oligonucleotide derived from the IgE germline promoter containing the PU.1 binding site and migrated with the same mobility compared with the complex formed with nuclear extracts. Transient transfection experiments using IgE germline promoter reporter gene constructs demonstrated that mutations affecting DNA binding of PU.1 or NF kappa B/rel had no or little effect on IL-4 inducibility of these plasmids. However, point mutations that abolished binding of both factors abrogated cytokine inducibility. No strict spacing of the STAT6 and the composite PU. 1/NF-kappa B elements is required for IL-4 induction. IL-4-induced STAT6 DNA binding was retained in PU.1-/NF kappa B/rel- double mutants. The data demonstrate that cooperation of STAT6 with at least PU.1 or NF kappa B/rel is necessary for IL-4-induced activation of IgE germline gene transcription.
Stütz
A M
AM
Department of Immunology, Novartis Research Institute, Vienna, Austria.
Woisetschläger
M
M
D000276:Adjuvants, Immunologic / Q000378:metabolism / Q000502:physiology*; D001483:Base Sequence; D004268:DNA-Binding Proteins / Q000502:physiology; D004357:Drug Synergism; D005786:Gene Expression Regulation / Q000276:immunology*; D006801:Humans; D007073:Immunoglobulin E / Q000235:genetics* / Q000378:metabolism; D015847:Interleukin-4 / Q000096:biosynthesis / Q000235:genetics / Q000502:physiology*; D008969:Molecular Sequence Data; D016328:NF-kappa B / Q000378:metabolism / Q000502:physiology*; D011401:Promoter Regions, Genetic / Q000276:immunology; D011485:Protein Binding / Q000235:genetics / Q000276:immunology; D011518:Proto-Oncogene Proteins / Q000235:genetics / Q000378:metabolism / Q000502:physiology*; D050783:Proto-Oncogene Proteins c-ets; D050800:STAT6 Transcription Factor; D015534:Trans-Activators / Q000235:genetics / Q000378:metabolism / Q000502:physiology*; D014157:Transcription Factors / Q000235:genetics / Q000378:metabolism; D014158:Transcription, Genetic / Q000276:immunology*; D014407:Tumor Cells, Cultured; D000099232:Proto-Oncogene Protein Spi-1
D000276:Adjuvants, Immunologic; D004268:DNA-Binding Proteins; D007073:Immunoglobulin E; D015847:Interleukin-4; D016328:NF-kappa B; D011518:Proto-Oncogene Proteins; D050783:Proto-Oncogene Proteins c-ets; D050800:STAT6 Transcription Factor; D015534:Trans-Activators; D014157:Transcription Factors; D000099232:Proto-Oncogene Protein Spi-1; C494101:STAT6 protein, human