Isolation of the Cyclosporin-Sensitive T Cell Transcription Factor NFATp

• Published in: Science (American Association for the Advancement of Science) 1993-10, Vol.262 (5134), p.750-754
• Main Authors: McCaffrey, Patricia G., Luo, Chun, Kerppola, Tom K., Jain, Jugnu, Badalian, Tina M., Ho, Andrew M., Burgeon, Emmanuel, Lane, William S., Lambert, John N., Curran, Tom, Verdine, Gregory L., Rao, Anjana, Hogan, Patrick G.
• Format: Article
• Language: English
• Subjects: Amino Acid Sequence; Amino acids; Animals; Antiserum; Base Sequence; Biological and medical sciences; Cell Line; Complementary DNA; DNA; DNA, Complementary; DNA-Binding Proteins - genetics; DNA-Binding Proteins - isolation & purification; DNA-Binding Proteins - physiology; Fundamental and applied biological sciences. Psychology; Gels; Immunosuppressive Agents - pharmacology; Interleukin-2 - genetics; Mice; Molecular and cellular biology; Molecular genetics; Molecular Sequence Data; NFATC Transcription Factors; Nuclear Proteins; Oligonucleotides; Phosphoproteins - genetics; Phosphoproteins - isolation & purification; Phosphoproteins - physiology; Promoter Regions, Genetic; Proto-Oncogene Proteins c-fos - physiology; Proto-Oncogene Proteins c-jun - physiology; Recombinant Proteins; RNA, Messenger - analysis; T lymphocytes; T-Lymphocytes - chemistry; Transcription factors; Transcription Factors - genetics; Transcription Factors - isolation & purification; Transcription Factors - physiology; Transcription. Transcription factor. Splicing. Rna processing
• ISSN: 0036-8075; 1095-9203
• DOI: 10.1126/science.8235597

Nuclear factor of activated T cells (NFAT) is a transcription factor that regulates expression of the cytokine interleukin-2 (IL-2) in activated T cells. The DNA-binding specificity of NFAT is conferred by NFATp, a phosphoprotein that is a target for the immunosuppressive compounds cyclosporin A and FK506. Here, the purification of NFATp from murine T cells and the isolation of a complementary DNA clone encoding NFATp are reported. A truncated form of NFATp, expressed as a recombinant protein in bacteria, binds specifically to the NFAT site of the murine IL-2 promoter and forms a transcriptionally active complex with recombinant c-Fos and c-Jun. Antisera to tryptic peptides of the purified protein or to the recombinant protein fragment react with T cell NFATp. The molecular cloning of NFATp should allow detailed analysis of a T cell transcription factor that is central to initiation of the immune response.