Journal article
Identification of Novel Nuclear Factor of Activated T Cell (NFAT)-associated Proteins in T Cells



Publication Details
Authors:
Gabriel, C.; Gross, F.; Karl, M.; Stephanowitz, H.; Hennig, A.; Weber, M.; Gryzik, S.; Bachmann, I.; Hecklau, K.; Wienands, J.; Schuchhardt, J.; Herzel, H.; Radbruch, A.; Krause, E.; Baumgrass, R.
Publisher:
AMER SOC BIOCHEMISTRY MOLECULAR BIOLOGY INC
Publication year:
2016
Journal:
Journal of Biological Chemistry
Pages range:
24172-24187
Volume number:
291
Issue number:
46
Start page:
24172
End page:
24187
Number of pages:
16
ISSN:
0021-9258

Abstract
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/A, NFATc1/C, 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.


Keywords
AP1 transcription factor (AP1), cAMP-response element-binding protein (CREB), lymphocyte, mass spectrometry (MS), NFAT transcription factor, protein-protein interaction, RUNX transcription factor, SILAC

Last updated on 2019-25-07 at 19:29