Lymphoid enhancer factor interacts with GATA-3 and controls its function in T helper type 2 cells
Lymphoid enhancer factor interacts with GATA-3 and controls its function in T helper type 2 cells
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DOI：
10.1111/j.1365-2567.2008.02854.x
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发表时间：
2008-11-01
期刊：
影响因子：
6.4
通讯作者：
Nakayama, Toshinori
中科院分区：
文献类型：
--
作者：
Hossain, Mohammad B.;Hosokawa, Hiroyuki;Nakayama, Toshinori
GATA-3是Th2细胞分化的主要转录因子，对Th2细胞因子的表达起关键作用。然而，人们对GATA-3的功能分子络合物的性质知之甚少。我们利用基于Flag-钙调蛋白结合肽(CBP)标签的蛋白质组学方法，在Th2细胞的GATA-3复合体中鉴定了一个高迁移率族(HMG)盒型转录因子-淋巴增强因子1(Lef-1)。GATA-3与Lef-1的免疫共沉淀实验证实了GATA-3与Lef-1之间的相互作用。Lef-1的HMG-box结构域和GATA-3的两个锌指结构域对物理结合是重要的。在发育中的Th2细胞中引入Lef-1可抑制Th2细胞因子的产生。在引入HMG盒缺失的Lef-1突变体的细胞中，这种抑制作用明显较低。此外，Lef-1抑制GATA-3与IL-5启动子的结合活性。这些结果表明，Lef-1参与了GATA-3复合体，同时也调节了GATA-3的功能，如通过抑制GATA-3的DNA结合活性诱导Th2细胞因子的表达。
GATA-3 is the master transcription factor for T helper 2 (Th2) cell differentiation and is critical for the expression of Th2 cytokines. Little is known, however, about the nature of the functional molecular complexes of GATA-3. We identified a high-mobility group (HMG)-box type transcription factor, lymphoid enhancer factor 1 (LEF-1), in the GATA-3 complex present in Th2 cells using a Flag-calmodulin-binding peptide (CBP)-tag based proteomics method. The interaction between GATA-3 and LEF-1 was confirmed by co-immunoprecipitation experiments using LEF-1-introduced T-cell lineage TG40 cells. The HMG-box domain of LEF-1 and two zinc finger domains of GATA-3 were found to be important for the physical association. The introduction of LEF-1 into developing Th2 cells resulted in the suppression of Th2 cytokine production. The suppression was significantly lower in the cells into which a HMG-box-deleted LEF-1 mutant was introduced. Moreover, LEF-1 inhibited the binding activity of GATA-3 to the interleukin (IL)-5 promoter. These results suggest that LEF-1 is involved in the GATA-3 complex, while also regulating the GATA-3 function, such as the induction of Th2 cytokine expression via the inhibition of the DNA-binding activity of GATA-3.