A transcription factor with AP3-like binding specificity mediates gene regulation after an allergic triggering with IgE and Ag in mouse mast cells.
Mast cells are an important source of a number of lymphokines and chemokines primarily those released after challenge with the allergic trigger IgE and Ag. However, the mechanisms of lymphokine and chemokine gene activation in this cell type, as opposed to the mechanisms of activation in T cells, are poorly understood. As a model system, we addressed this issue in mast cells by using the recently cloned chemokine MARC gene, which belongs to the RANTES/sis gene family. After allergic stimulation, MARC induction is pronounced and mast cell specific. Northern blot analysis, in combination with two inhibitors, actinomycin D and cycloheximide, resulted in the formation of our initial hypothesis, which was that both transcriptional and post-transcriptional regulation are involved after stimulation through the Fc epsilon R. We performed a detailed promoter analysis of the cloned MARC gene by using transient assays of transfected reporter gene constructs. Thereby, two potential promoter regions were identified as being crucial for transcriptional stimulation. Additional fine mapping of the proximal element and subsequent electrophoretic mobility shift assays, combined with competitions of known transcription factor binding sites, identified one of the transcription factors in stimulated mast cells as an AP3 or AP3-like binding activity.
Jarmin
D I
DI
Department of Immunoregulation, Sandoz Research Institute, Vienna, Austria.
Kulmburg
P A
PA
Huber
N E
NE
Baumann
G
G
Prieschl-Strassmayr
E E
EE
Baumruker
T
T
D000595:Amino Acid Sequence; D000818:Animals; D000941:Antigens / Q000276:immunology*; D001483:Base Sequence; D054410:Chemokine CCL7; D003001:Cloning, Molecular; D016207:Cytokines / Q000096:biosynthesis / Q000235:genetics*; D004268:DNA-Binding Proteins / Q000378:metabolism; D005786:Gene Expression Regulation / Q000502:physiology*; D015698:Genomic Library; D006967:Hypersensitivity / Q000276:immunology; D007073:Immunoglobulin E / Q000276:immunology*; D008407:Mast Cells / Q000502:physiology; D051379:Mice; D008969:Molecular Sequence Data; D018945:Monocyte Chemoattractant Proteins; D011401:Promoter Regions, Genetic / Q000235:genetics; D012045:Regulatory Sequences, Nucleic Acid / Q000502:physiology; D014157:Transcription Factors / Q000378:metabolism / Q000502:physiology*; D014158:Transcription, Genetic / Q000235:genetics; D015533:Transcriptional Activation
D000941:Antigens; C515451:Ccl7 protein, mouse; D054410:Chemokine CCL7; D016207:Cytokines; D004268:DNA-Binding Proteins; D018945:Monocyte Chemoattractant Proteins; D014157:Transcription Factors; C060759:enhancer-binding protein AP-3; D007073:Immunoglobulin E