Genomic code for Sox10 activation reveals a key regulatory enhancer for cranial neural crest
Genomic code for Sox10 activation reveals a key regulatory enhancer for cranial neural crest
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DOI：
10.1073/pnas.0906596107
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发表时间：
2010-02-23
影响因子：
11.1
通讯作者：
Sauka-Spengler, Tatjana
中科院分区：
文献类型：
--
作者：
Betancur, Paola;Bronner-Fraser, Marianne;Sauka-Spengler, Tatjana
神经嵴是一种多能的干细胞样细胞群，在胚胎中广泛迁移并形成广泛的衍生物，从神经元到黑素细胞和软骨。对驱动神经嵴发育的基因调控网络的分析表明，Sox 10是最早的神经嵴特异性基因之一，细胞自主驱动分层并直接调控许多下游效应子和分化基因电池。在寻找直接输入到神经嵴指定模块，我们解剖了鸡Sox10基因组区域，并分离出两个下游调控区域具有不同的时空活动。一个独特的元素，Sox10E2代表最早作用的神经嵴顺式调节元件，启动Sox10表达在新形成的颅，但不是迷走神经和躯干神经嵴的关键。第二个元素，Sox10E1，在后来迁移迷走神经和躯干嵴细胞的行为。Sox10E2的深入表征揭示了Sox9，Ets1和cMyb作为介导增强子活性的直接输入。ChIP，DNA下拉，凝胶转移试验表明，它们直接结合到体内的Sox10E2增强子，而其相应的结合位点的突变，或失活的三个上游监管机构，废除报告和内源性Sox10的表达。使用顺式调控分析作为工具，我们的研究在神经嵴基因调控网络中建立了关键的连接，从而在建立上游效应子与关键神经嵴指定符的直接联系方面是独一无二的。
The neural crest is a multipotent, stem cell-like population that migrates extensively in the embryo and forms a wide array of derivatives, ranging from neurons to melanocytes and cartilage. Analyses of the gene regulatory network driving neural crest development revealed Sox10 as one of the earliest neural crest-specifying genes, cell-autonomously driving delamination and directly regulating numerous downstream effectors and differentiation gene batteries. In search of direct inputs to the neural crest specifier module, we dissected the chick Sox10 genomic region and isolated two downstream regulatory regions with distinct spatiotemporal activity. A unique element, Sox10E2 represents the earliest-acting neural crest cis-regulatory element, critical for initiating Sox10 expression in newly formed cranial, but not vagal and trunk neural crest. A second element, Sox10E1, acts in later migrating vagal and trunk crest cells. Deep characterization of Sox10E2 reveals Sox9, Ets1, and cMyb as direct inputs mediating enhancer activity. ChIP, DNA-pull down, and gel-shift assays demonstrate their direct binding to the Sox10E2 enhancer in vivo, whereas mutation of their corresponding binding sites, or inactivation of the three upstream regulators, abolishes both reporter and endogenous Sox10 expression. Using cis-regulatory analysis as a tool, our study makes critical connections within the neural crest gene regulatory network, thus being unique in establishing a direct link of upstream effectors to a key neural crest specifier.