Regulation of expression of the IL-2 and IL-5 genes and the role of proteins related to nuclear factor of activated T cells.
Cyclic adenosine monophosphate (cAMP) inhibits phorbol 12-myristate 13-acetate (PMA)-induced IL-2 production while it inhibits IL-5 production at the transcriptional level in EL-4, a mouse lymphoma line. The -321 to +46 region of the mouse IL-2 promoter is required for activation by PMA and inhibition by cAMP. This promoter region contains several elements that interact with transcription factors, such as nuclear factor of activated T cells (NF-AT), NF-kappa B, AP-1, and octamer. With use of reporter plasmid carrying multiple copies of each element, we found that the construct that contained the NF-AT site was most effective for responding to PMA activation and cAMP inhibition. In electrophoretic mobility shift assay (EMSA), PMA-induced NF-AT binding complex was altered by cAMP. Furthermore, overexpression of the cytoplasmic component of NF-AT abrogated the inhibitory action of cAMP. These results indicate that the NF-AT site is a target of the inhibitory action of cAMP. We have previously reported that the -1200 to +33 region of the mouse IL-5 promoter can mediate transcriptional stimulation by PMA and cAMP in EL-4 cells. Here we identified the element IL-5P, which is required for maximal activation of the IL-5 promoter. We found that this element is homologous to the binding site for NF-AT and interacted with NF-AT-related factors induced by PMA and cAMP. Thus it appears that an NF-AT factor is involved in the regulation of IL-5 gene transcription.
Tsuruta
L
L
Department of Molecular and Developmental Biology, University of Tokyo, Japan.
Lee
H J
HJ
Masuda
E S
ES
Yokota
T
T
Arai
N
N
Arai
K
K
D000818:Animals; D001483:Base Sequence; D001665:Binding Sites; D000242:Cyclic AMP / Q000494:pharmacology*; D018076:DNA, Complementary / Q000235:genetics; D004268:DNA-Binding Proteins / Q000235:genetics / Q000502:physiology*; D005786:Gene Expression Regulation / Q000502:physiology*; D007376:Interleukin-2 / Q000096:biosynthesis / Q000235:genetics*; D015848:Interleukin-5 / Q000096:biosynthesis / Q000235:genetics*; D008213:Lymphocyte Activation / Q000502:physiology*; D008223:Lymphoma / Q000235:genetics / Q000378:metabolism; D051379:Mice; D008969:Molecular Sequence Data; D050778:NFATC Transcription Factors; D009687:Nuclear Proteins / Q000235:genetics / Q000502:physiology*; D011401:Promoter Regions, Genetic / Q000502:physiology; D012689:Sequence Homology, Nucleic Acid; D013601:T-Lymphocytes / Q000502:physiology*; D013755:Tetradecanoylphorbol Acetate / Q000037:antagonists & inhibitors / Q000494:pharmacology; D014157:Transcription Factors / Q000235:genetics / Q000502:physiology*; D014158:Transcription, Genetic / Q000187:drug effects; D014162:Transfection; D014407:Tumor Cells, Cultured
D018076:DNA, Complementary; D004268:DNA-Binding Proteins; D007376:Interleukin-2; D015848:Interleukin-5; D050778:NFATC Transcription Factors; D009687:Nuclear Proteins; D014157:Transcription Factors; D000242:Cyclic AMP; D013755:Tetradecanoylphorbol Acetate