DLX3 interacts with GCM1 and inhibits its transactivation-stimulating activity in a homeodomain-dependent manner in human trophoblast-derived cells
DLX3 interacts with GCM1 and inhibits its transactivation-stimulating activity in a homeodomain-dependent manner in human trophoblast-derived cells
复制标题
DOI：
10.1038/s41598-017-02120-5
复制
发表时间：
2017-05-17
影响因子：
4.6
通讯作者：
Roberson, Mark S.
中科院分区：
文献类型：
--
作者：
Li, Sha;Roberson, Mark S.
胎盘转录因子Distal-less 3(DLX3)和胶质细胞缺失-1(GCM1)在人滋养层细胞系中协调PGF的特异性调控。虽然这两个因素单独对PGF基因的表达有积极影响，但当两者联合使用时，DLX3对GCM具有拮抗剂的作用。尽管有了这样的理解，但解释这种监管相互作用的潜在机制仍未被探索。我们通过进行免疫沉淀和哺乳动物单杂交实验，鉴定了人滋养层细胞中DLX3和GCM1的特定结构域之间的物理和功能相互作用。研究表明，DLX3结合降低了GCM1的转录活性，为其调节PGF启动子活性的功能拮抗提供了机制解释。DLX3同源结构域(HD)是DLX3-GCM1相互作用所必需的，而HD与DLX3的氨基或羧基末端结构域一起是最大限度地抑制GCM1所必需的。有趣的是，在这个系统中，一个自然产生的DLX3突变体破坏了导致人类毛发齿骨综合征的羧基末端结构域，表现出与野生型DLX3无法区分的活动。综上所述，我们的研究表明DLX3与GCM1在物理上相互作用并抑制其反式激活活性，提示DLX3和GCM1可能形成一个复合体，通过调节靶基因的表达来调节胎盘细胞的功能。
The placental transcription factors Distal-less 3 (DLX3) and Glial cell missing-1 (GCM1) have been shown to coordinate the specific regulation of PGF in human trophoblast cell lines. While both factors independently have a positive effect on PGF gene expression, when combined, DLX3 acts as an antagonist to GCM. Despite this understanding, potential mechanisms accounting for this regulatory interaction remain unexplored. We identify physical and functional interactions between specific domains of DLX3 and GCM1 in human trophoblast-derived cells by performing immunoprecipitation and mammalian one hybrid assays. Studies revealed that DLX3 binding reduced the transcriptional activity of GCM1, providing a mechanistic explanation of their functional antagonism in regulating PGF promoter activity. The DLX3 homeodomain (HD) was essential for DLX3-GCM1 interaction, and that the HD together with the DLX3 amino-or carboxyl-terminal domains was required for maximal inhibition of GCM1. Interestingly, a naturally occurring DLX3 mutant that disrupts the carboxyl-terminal domain leading to tricho-dento-osseous syndrome in humans displayed activities indistinguishable from wild type DLX3 in this system. Collectively, our studies demonstrate that DLX3 physically interacts with GCM1 and inhibits its transactivation activity, suggesting that DLX3 and GCM1 may form a complex to functionally regulate placental cell function through modulation of target gene expression.