Abstract
Four nuclear factor of activated T cells (NF-AT) binding motifs were found in the murine CD40 ligand promoter. Electrophoretic mobility shift assays using 18-base pair (bp) long oligonucleotides corresponding to the proximal site and nuclear extracts from activated T cells revealed two complexes which were inhibited by cyclosporin A and contained NF-ATc and NF-ATp. Neither complex contained AP-1 proteins. Multimers of the 18-bp oligonucleotides were not active in transient transfection assays using luciferase reporter gene constructs. In contrast, a 30-bp long oligonucleotide bound AP-1 proteins in addition to NF-AT proteins and its multimers strongly induced luciferase gene expression. These results suggested that NF-AT proteins play an important role in the expression of the CD40L gene and that their transcriptional activity requires AP-1 binding.
Publication types
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Research Support, Non-U.S. Gov't
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Research Support, U.S. Gov't, P.H.S.
MeSH terms
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Animals
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Base Sequence
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Binding Sites
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CD40 Antigens / immunology
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CD40 Ligand
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Cell Line
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Cell Nucleus / metabolism
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Consensus Sequence
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DNA-Binding Proteins / metabolism*
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Flow Cytometry
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Interleukin-2 / genetics
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Luciferases / biosynthesis
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Lymphocyte Activation
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Membrane Glycoproteins / biosynthesis*
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Membrane Glycoproteins / genetics*
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Mice
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Molecular Sequence Data
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NFATC Transcription Factors
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Nuclear Proteins / metabolism
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Oligodeoxyribonucleotides
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Promoter Regions, Genetic*
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Recombinant Proteins / biosynthesis
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T-Lymphocytes / immunology*
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T-Lymphocytes / metabolism
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Transcription Factor AP-1 / metabolism*
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Transcription Factors / metabolism*
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Transcriptional Activation*
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Tumor Cells, Cultured
Substances
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CD40 Antigens
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DNA-Binding Proteins
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Interleukin-2
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Membrane Glycoproteins
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NFATC Transcription Factors
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Nuclear Proteins
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Oligodeoxyribonucleotides
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Recombinant Proteins
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Transcription Factor AP-1
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Transcription Factors
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CD40 Ligand
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Luciferases