Identification of Novel Nuclear Factor of Activated T Cell (NFAT)-associated Proteins in T Cells
Identification of Novel Nuclear Factor of Activated T Cell (NFAT)-associated Proteins in T Cells
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DOI：
10.1074/jbc.m116.739326
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发表时间：
2016-11-11
影响因子：
4.8
通讯作者：
Baumgrass, Ria
中科院分区：
文献类型：
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作者：
Gabriel, Christian H.;Gross, Fridolin;Baumgrass, Ria
活化T细胞核因子（NFAT）家族的转录因子是抗原特异性T细胞活化和分化所必需的。它们与其他转录因子如AP 1蛋白（FOS、JUN和JUNB）、FOXP 3、IRFs和EGR 1的协同DNA结合决定了NFAT的基因调控作用。为了鉴定NFAT的未知相互作用伙伴，我们纯化了生物素标记的NFATc 1/A、NFATc 1/C和NFATc 2/C蛋白复合物，并通过基于细胞培养的质谱中的氨基酸稳定同位素标记来分析它们的组分。我们发现了超过170个NFAT相关蛋白，其中一半参与转录调控。其中有许多迄今未知的NFATc 1和NFATc 2在T细胞中的相互作用伴侣，如Raptor、CHEK 1、CREB 1、RUNX 1、SATB 1、Ikaros和Helios。NFATc 2与其他几种转录因子的关联是DNA依赖性的，表明协同DNA结合。此外，我们的计算分析发现，RUNX和CREB 1的结合基序被发现优先在NFAT结合基序的直接附近，并在一个不同的方向。此外，我们提供了mTOR和CHEK 1激酶活性影响NFAT转录效力的证据。最后，我们的NFAT相关蛋白的数据集为进一步研究NFAT的多种功能以及这些功能如何由于多个相互作用伙伴的相互作用而被调节提供了良好的基础。
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.