Cloning and characterization of the cardiac-specific Lrrc10 promoter
Cloning and characterization of the cardiac-specific Lrrc10 promoter
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心脏特异性 Lrrc10 启动子的克隆和表征
DOI：
10.5483/bmbrep.2011.44.2.123
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发表时间：
2011-02-28
期刊：
影响因子：
3.8
通讯作者：
Wu, Xiushan
中科院分区：
文献类型：
--
作者：
Fan, Xiongwei;Yang, Qing;Wu, Xiushan
富含亮氨酸重复序列的蛋白质10（LRRC 10）是一个心脏特异性基因，在心脏发育和疾病中发挥作用。最近报道了斑马鱼morphants中严重的心脏形态发生缺陷，但在小鼠中发现了矛盾的结果，表明小鼠胚胎发育过程中存在更复杂的分子机制。为了阐明LRRC 10是如何被调节的，我们使用荧光素酶报告基因测定法分析了Lrrc 10起始位点上游约3千碱基（kb）的5 '增强子区域。我们对Lrrc 10启动子的表征表明它具有复杂的顺式和反式作用元件。我们发现，GATA 4和MEF 2C都可以单独增加Lrrc 10启动子的转录活性，但它们并不协同，这表明存在更复杂的调控模式。令人惊讶的是，敲除5 '增强子区域中的Gata 4和Mef 2c结合位点（-2，894/-2，889）不改变Lrrc 10启动子的转录活性，并且鉴定的可能的GATA 4结合位点位于启动子上游仅100个碱基对（bp）的区域中。我们的数据提供了深入了解Lrrc 10表达的分子调控，这可能也有助于其组织特异性表达。[BMB报告2011; 44（2）：123-128]
Leucine-rich repeat containing protein 10 (LRRC10) is characterized as a cardiac-specific gene, suggesting a role in heart development and disease. A severe cardiac morphogenic defect in zebrafish morphants was recently reported but a contradictory result was found in mice, suggesting a more complicated molecular mechanism exists during mouse embryonic development. To elucidate how LRRC10 is regulated, we analyzed the 5'enhancer region approximately 3 kilo bases (kb) upstream of the Lrrc10 start site using luciferase reporter gene assays. Our characterization of the Lrrc10 promoter indicates it possesses complicated cis-and trans-acting elements. We show that GATA4 and MEF2C could both increase transcriptional activity of Lrrc10 promoter individually but that they do not ad synergistically, suggesting that there exists a more complex regulation pattern. Surprisingly, knockout of Gata4 and Mef2c binding sites in the 5'enhancer region (-2,894/-2,889) didn't change the transcriptional activity of the Lrrc10 promoter and the likely GATA4 binding site identified was located in a region only 100 base pair (bp) upstream of the promoter. Our data provides insight into the molecular regulation of Lrrc10 expression, which probably also contributes to its tissue-specific expression. [BMB reports 2011; 44(2): 123-128]