Identification of Novel Nuclear Factor of Activated T Cell (NFAT)-associated Proteins in T Cells
| Autor: | Stefanie Gryzik, Jürgen Wienands, Martin Karl, Melanie Weber, Johannes Schuchhardt, Katharina Hecklau, Fridolin Gross, Ivo Bachmann, Ria Baumgrass, Hanspeter Herzel, Heike Stephanowitz, Christian H. Gabriel, Eberhard Krause, Anna Floriane Hennig, Andreas Radbruch |
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| Rok vydání: | 2016 |
| Předmět: |
0301 basic medicine
JUNB T-Lymphocytes T cell Biology Biochemistry Mass Spectrometry Jurkat Cells 03 medical and health sciences 0302 clinical medicine Stable isotope labeling by amino acids in cell culture medicine Transcriptional regulation Humans Kinase activity Molecular Biology Transcription factor NFAT transcription factor RUNX transcription factor SILAC lymphocyte mass spectrometry (MS) protein-protein interaction Bioinformatics AP1 transcription factor (AP1) cAMP-response element-binding protein (CREB) NFATC Transcription Factors Nuclear Proteins NFAT Cell Biology Molecular biology Cell biology AP-1 transcription factor 030104 developmental biology medicine.anatomical_structure 030215 immunology |
| Zdroj: | Journal of applied biological chemistry, 291(46), 24172-24187 |
| ISSN: | 0021-9258 |
| Popis: | Transcription factors of the nuclear factor of activated T cell (NFAT) family are essential for antigen-specific T cell activation and differentiation. Their cooperative DNA binding with other transcription factors, such as AP1 proteins (FOS, JUN, and JUNB), FOXP3, IRFs, and EGR1, dictates the gene regulatory action of NFATs. To identify as yet unknown interaction partners of NFAT, we purified biotin-tagged NFATc1/αA, NFATc1/βC, and NFATc2/C protein complexes and analyzed their components by stable isotope labeling by amino acids in cell culture-based mass spectrometry. We revealed more than 170 NFAT-associated proteins, half of which are involved in transcriptional regulation. Among them are many hitherto unknown interaction partners of NFATc1 and NFATc2 in T cells, such as Raptor, CHEK1, CREB1, RUNX1, SATB1, Ikaros, and Helios. The association of NFATc2 with several other transcription factors is DNA-dependent, indicating cooperative DNA binding. Moreover, our computational analysis discovered that binding motifs for RUNX and CREB1 are found preferentially in the direct vicinity of NFAT-binding motifs and in a distinct orientation to them. Furthermore, we provide evidence that mTOR and CHEK1 kinase activity influence NFAT's transcriptional potency. Finally, our dataset of NFAT-associated proteins provides a good basis to further study NFAT's diverse functions and how these are modulated due to the interplay of multiple interaction partners. |
| Databáze: | OpenAIRE |
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