Identification of novel interacting regions involving calcineurin and nuclear factor of activated T cells
Identification of novel interacting regions involving calcineurin and nuclear factor of activated T cells
复制标题
DOI：
10.1096/fj.201902229
复制
发表时间：
2020-01
期刊：
影响因子：
--
通讯作者：
N. Kitamura;M. Shindo;J. Ohtsuka;A. Nakamura;M. Tanokura;T. Hiroi;O. Kaminuma
中科院分区：
文献类型：
--
作者：
N. Kitamura;M. Shindo;J. Ohtsuka;A. Nakamura;M. Tanokura;T. Hiroi;O. Kaminuma
活化T细胞核因子(NFAT)可导致多种可诱导基因的转录，参与多种生物学过程，因此，NFAT的异常表达与多种疾病的发生和发展密切相关。由于NFAT的五种异构体(NFATc1-c4，NFAT5)具有不同且重叠的功能，因此需要选择性地控制NFAT家族成员的一部分，而不是全部。通过比较每个NFATc1-c4与其调控酶钙调神经磷酸酶(CN)的结合活性，我们发现在NFATc1和NFATc4中有一个新的CN结合区域(CNBR)选择性地发挥作用。这个区域被称为CNBR3，位于钙离子调节域中两个先前存在的CNBR1和CNBR2之间。CNBR3的异位表达抑制了NFATc1而不是NFATc2在T细胞中的核转位，从而下调了NFATc1依赖的细胞因子的表达。通过NFATc1部分肽的竞争分析和光亲和质谱分析，我们鉴定了NFATc1的18个氨基酸(Arg258~Pro275)和CN催化亚基(Asn77~Gly89)的13个氨基酸负责CNA/CNBR3的结合，其中Cys263和Asp82分别在其中起关键作用。建议通过靶向这个新的CN/NFAT结合区来选择性地调节NFAT介导的生物过程。
Nuclear factor of activated T cells (NFAT) leads to the transcription of diverse inducible genes involved in many biological processes; therefore, aberrant NFAT expression is responsible for the development and exacerbation of various disorders. Since five isoforms of NFAT (NFATc1‐c4, NFAT5) exhibit distinct and overlapping functions, selective control of a part, but not all, of NFAT family members is desirable. By comparing the binding activity of each NFATc1‐c4 with its regulatory enzyme, calcineurin (CN), using a quantitative immunoprecipitation assay, we found a new CN‐binding region (CNBR) selectively functioning in NFATc1 and NFATc4. This region, termed CNBR3, is located between two preexisting CNBR1 and CNBR2, within the Ca2+ regulatory domain. The nuclear translocation of NFATc1 but not NFATc2 in T cells was suppressed by ectopic expression of CNBR3 and, accordingly, NFATc1‐dependent cytokine expression was downregulated. Through competition assays using NFATc1‐derived partial peptides and mass spectrometry with photoaffinity technology, we identified 18 amino acids in NFATc1 (Arg258 to Pro275) and 13 amino acids in CN catalytic subunit (CNA) (Asn77 to Gly89) responsible for CNA/CNBR3 binding in which Cys263 and Asp82, respectively, played crucial roles. The possible selective regulation of NFAT‐mediated biological processes by targeting this new CN/NFAT‐binding region is suggested.