SOX10 directly modulates ERBB3 transcription via an intronic neural crest enhancer.
SOX10 directly modulates ERBB3 transcription via an intronic neural crest enhancer.
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DOI：
10.1186/1471-213x-11-40
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发表时间：
2011-06-14
影响因子：
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通讯作者：
McCallion AS
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文献类型：
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作者：
Prasad MK;Reed X;Gorkin DU;Cronin JC;McAdow AR;Chain K;Hodonsky CJ;Jones EA;Svaren J;Antonellis A;Johnson SL;Loftus SK;Pavan WJ;McCallion AS
ERBB3基因对于神经脊(NC)及其衍生群体(如雪旺细胞)的正常发育至关重要。与所有细胞命运的决定一样，转录调控在发育过程中对NC来源的谱系进行限制和规范方面发挥着重要作用。然而，对介导ERBB3转录调控的序列或结合它们的因素知之甚少。在这项研究中，我们鉴定了ERBB3基因上的三个转录增强子，并在体外对NC来源的细胞类型和在体内对转基因斑马鱼的调控潜力进行了评估。一种称为ERBB3_MCS6的增强子位于ERBB3的第一内含子中，它在体外引导最高的报告基因表达，并显示出与增强子活性一致的表观遗传标记。我们在ERBB3_MCS6中确定了一个共识的Sox10结合位点，并在体外证明了它对该增强子活性的必要性和充分性。此外，我们还证明了内源性erbB3基因的转录依赖于Sox10。进一步，我们在体外证明了Sox10与ERBB3_MCS6在物理上相互作用。与其体外活性一致，我们还发现ERBB3_MCS6以一种与erb3b表达一致的方式在NC细胞及其在斑马鱼体内的衍生谱系中驱动报告基因的表达。我们还利用Morolino分析证明了Sox10是ERBB3_MCS6在斑马鱼体内表达所必需的。综上所述，我们的数据表明，ERBB3可能受到Sox10的直接调控，这种调控可能在一定程度上受到ERBB3_MCS6的促进。
The ERBB3 gene is essential for the proper development of the neural crest (NC) and its derivative populations such as Schwann cells. As with all cell fate decisions, transcriptional regulatory control plays a significant role in the progressive restriction and specification of NC derived lineages during development. However, little is known about the sequences mediating transcriptional regulation of ERBB3 or the factors that bind them. In this study we identified three transcriptional enhancers at the ERBB3 locus and evaluated their regulatory potential in vitro in NC-derived cell types and in vivo in transgenic zebrafish. One enhancer, termed ERBB3_MCS6, which lies within the first intron of ERBB3, directs the highest reporter expression in vitro and also demonstrates epigenetic marks consistent with enhancer activity. We identify a consensus SOX10 binding site within ERBB3_MCS6 and demonstrate, in vitro, its necessity and sufficiency for the activity of this enhancer. Additionally, we demonstrate that transcription from the endogenous Erbb3 locus is dependent on Sox10. Further we demonstrate in vitro that Sox10 physically interacts with that ERBB3_MCS6. Consistent with its in vitro activity, we also show that ERBB3_MCS6 drives reporter expression in NC cells and a subset of its derivative lineages in vivo in zebrafish in a manner consistent with erbb3b expression. We also demonstrate, using morpholino analysis, that Sox10 is necessary for ERBB3_MCS6 expression in vivo in zebrafish. Taken collectively, our data suggest that ERBB3 may be directly regulated by SOX10, and that this control may in part be facilitated by ERBB3_MCS6.