Activation of the interleukin-4 gene is controlled by the unique calcineurin-dependent transcriptional factor NF(P).
Cyclosporin A-sensitive promoter activity that is induced by the T cell antigen receptor was identified in the region between base pairs -147 and -17 of the mouse interleukin (IL)-4 gene. Here, we identify a transcriptional factor, NF(P), that binds a sequence from -75 to -69 (ATTTTCC) of the mouse IL-4 gene which corresponds to the human IL-4 gene P sequence. Expression of NF(P) was associated with positive regulation of the IL-4 gene and cyclosporin A sensitivity. The P sequence is homologous to the binding site for the positive transcriptional regulator of the IL-2 gene nuclear factor of activated T cells (NFAT). Binding of NF(P) to the P sequence was completely inhibited by an oligonucleotide corresponding to the NFAT binding site. Like NFAT, NF(P) was also found in the cytosol of resting T cells but translocated to the nucleus via a cyclosporin A-sensitive mechanism after T cell activation. Overexpression of a constitutively active calcineurin caused translocation of NF(P) from the cytosol to the nucleus. NF(P) purified on an oligonucleotide affinity column was 90 kDa, which is smaller than the 120-kDa reported for cNFAT. Purified NF(P) did not bind the NFAT consensus sequence, even in the presence of AP-1 protein. Our results suggest that the NF(P) molecule is not identical to the cNFAT molecule.
Department of Inflammation Biology and Immunology, Syntex Discovery Research, Palo Alto, California 94304.
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M
Kubo
R L
RL
Kincaid
J T
JT
Ransom
D000818:Animals; D001483:Base Sequence; D019703:Calcineurin; D002118:Calcium / Q000378:metabolism; D002148:Calmodulin-Binding Proteins / Q000378:metabolism*; D002451:Cell Compartmentation; D002467:Cell Nucleus / Q000378:metabolism; D016384:Consensus Sequence; D016572:Cyclosporine / Q000494:pharmacology; D003600:Cytosol / Q000737:chemistry; D004268:DNA-Binding Proteins / Q000378:metabolism; D007376:Interleukin-2 / Q000378:metabolism; D015847:Interleukin-4 / Q000235:genetics*; D051379:Mice; D008969:Molecular Sequence Data; D050778:NFATC Transcription Factors; D009687:Nuclear Proteins; D010749:Phosphoprotein Phosphatases / Q000378:metabolism*; D011401:Promoter Regions, Genetic / Q000235:genetics*; D015398:Signal Transduction / Q000187:drug effects; D014157:Transcription Factors / Q000378:metabolism*
D002148:Calmodulin-Binding Proteins; D004268:DNA-Binding Proteins; D007376:Interleukin-2; D050778:NFATC Transcription Factors; D009687:Nuclear Proteins; D014157:Transcription Factors; D015847:Interleukin-4; D016572:Cyclosporine; D019703:Calcineurin; D010749:Phosphoprotein Phosphatases; D002118:Calcium