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首页> 外文期刊>BMC Molecular Biology >Ets-2 and C/EBP-beta are important mediators of ovine trophoblast Kunitz domain protein-1 gene expression in trophoblast
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Ets-2 and C/EBP-beta are important mediators of ovine trophoblast Kunitz domain protein-1 gene expression in trophoblast

机译：Ets-2和C / EBP-beta是绵羊滋养层中重要的介质，Kunitz域蛋白1基因在滋养层中的表达

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Background: The trophoblast Kunitz domain proteins (TKDPs) constitute a highly expressed, placenta-specific, multigene family restricted to ruminant ungulates and characterized by a C- terminal "Kunitz" domain, preceded by one or more unique N-terminal domains. TKDP-1 shares an almost identical expression pattern with interferon-tau, the "maternal recognition of pregnancy protein" in ruminants. Our goal here has been to determine whether the ovine (ov) Tkdp-1 and IFNT genes possess a similar transcriptional code. Results: The ovTkdp-1 promoter has been cloned and characterized. As with the IFNT promoter, the Tkdp-1 promoter is responsive to Ets-2, and promoter-driven reporter activity can be increased over 700-fold in response to over-expression of Ets-2 and a constitutively active form of protein Kinase A (PKA). Unexpectedly, the promoter element of Tkdp-1 responsible for this up-regulation, unlike that of the IFNT, does not bind Ets-2. However, mutation of a CCAAT/enhancer binding element within this control region not only reduced basal transcriptional activity, but prevented Ets-2 as well as cyclic adenosine 5'-monophosphate (cAMP)/PKA and Ras/mitogen-activated protein kinase (MAPK) responsiveness. In vitro binding experiments and in vivo protein-protein interaction assays implicated CCAAT/enhancer binding protein-beta (C/EBP-beta) as involved in up-regulating the Tkdp-1 promoter activity. A combination of Ets-2 and C/EBP-beta can up-regulate expression of the minimal Tkdp-1 promoter as much as 930-fold in presence of a cAMP analog. An AP-1-like element adjacent to the CCAAT enhancer, which binds Jun family members, is required for basal and cAMP/ C/EBP-β-dependent activation of the gene, but not for Ets-2-dependent activity. Conclusion: This paper demonstrates how Ets-2, a key transcription factor for trophoblast differentiation and function, can control expression of two genes (Tkdp-1 and IFNT) having similar spatial and temporal expression patterns via very different mechanisms.
机译：背景：滋养层的Kunitz结构域蛋白（TKDP）构成了一种高度表达的，胎盘特异性，多基因家族，只限于反刍动物有蹄类动物，并具有一个C端“ Kunitz”结构域，其后是一个或多个独特的N端结构域。 TKDP-1与反刍动物中的“妊娠蛋白的母体识别”干扰素-tau具有几乎相同的表达模式。我们的目标是确定绵羊（ov）Tkdp-1和IFNT基因是否具有相似的转录密码。结果：ovTkdp-1启动子已被克隆并鉴定。与IFNτ启动子一样，Tkdp-1启动子对Ets-2有反应，并且启动子驱动的报告子活性可以响应Ets-2的过表达和蛋白激酶A的组成型活性而增加700倍以上。 （PKA）。出乎意料的是，负责该上调的Tkdp-1的启动子元件与IFNτ的启动子元件不结合Ets-2。但是，在此控制区域内CCAAT /增强子结合元件的突变不仅降低了基础转录活性，而且阻止了Ets-2以及环状腺苷5'-单磷酸（cAMP）/ PKA和Ras /促分裂原活化蛋白激酶（MAPK） ）的响应能力。体外结合实验和体内蛋白质-蛋白质相互作用测定暗示CCAAT /增强子结合蛋白-β（C /EBP-β）与上调Tkdp-1启动子活性有关。在cAMP类似物的存在下，Ets-2和C / EBP-beta的组合可以上调最小Tkdp-1启动子的表达多达930倍。与CCAAT增强子相邻并结合Jun家族成员的AP-1样元件是该基因的基础和cAMP / C /EBP-β依赖性激活所必需的，而不是Ets-2依赖性活性所必需的。结论：本文证明了Ets-2是滋养细胞分化和功能的关键转录因子，如何通过非常不同的机制控制具有相似时空表达模式的两个基因（Tkdp-1和IFNτ）的表达。

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