CtBP2 Downregulation during Neural Crest Specification Induces Expression of Mitf and REST, Resulting in Melanocyte Differentiation and Sympathoadrenal Lineage Suppression
CtBP2 Downregulation during Neural Crest Specification Induces Expression of Mitf and REST, Resulting in Melanocyte Differentiation and Sympathoadrenal Lineage Suppression
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DOI：
10.1128/mcb.01062-10
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发表时间：
2011-03-01
影响因子：
5.3
通讯作者：
Andrisani, Ourania M.
中科院分区：
文献类型：
--
作者：
Liang, Hongzi;Fekete, Donna M.;Andrisani, Ourania M.
躯干神经嵴 (NC) 细胞分化为神经元、黑素细胞和神经胶质细胞。在 NC 培养物中，环磷酸腺苷 (cAMP) 会诱导黑素细胞分化，同时抑制神经元交感肾上腺谱系，具体取决于信号强度。黑素细胞分化需要激活 CREB 和 cAMP 依赖性蛋白激酶 A (PKA)，但 PKA 的作用尚不清楚。我们已经证明，在 NC 培养物中，cAMP 诱导小眼相关转录因子基因 (Mitf) 和 RE-1 沉默转录因子基因 (REST) 的转录，这两种基因都是 Wnt 调节的基因。在 NC 培养物和斑马鱼中，敲除 Wnt 介导的转录 C 端结合蛋白 2 (CtBP2) 的辅阻遏物（而非 CtBP1）可解除对 Mitf 和 REST 表达的抑制，并增强黑素细胞的分化。 NC 和 B16 黑色素瘤细胞中的 cAMP 降低了 CtBP2 蛋白水平，而抑制 PKA 或蛋白酶体则挽救了 CtBP2 降解。有趣的是，敲低同源结构域相互作用蛋白激酶 2 (HIPK2)（一种 CtBP 稳定性调节剂）可增加 CtBP2 水平，抑制 Mitf、REST 和黑素细胞分化的表达，并增加神经元基因表达和交感肾上腺谱系分化。我们得出结论，cAMP/PKA 通过 HIPK2 促进 CtBP2 降解，从而导致 Mitf 和 REST 表达。 Mitf 诱导黑素细胞分化，REST 抑制神经元特异性基因表达和交感肾上腺谱系。我们的研究确定了 REST 在 NC 细胞分化中的新作用，并表明在 NC 谱系规范中 cAMP 和 Wnt 信号传导之间存在交互作用。
Trunk neural crest (NC) cells differentiate to neurons, melanocytes, and glia. In NC cultures, cyclic AMP (cAMP) induces melanocyte differentiation while suppressing the neuronal sympathoadrenal lineage, depending on the signal intensity. Melanocyte differentiation requires activation of CREB and cAMP-dependent protein kinase A (PKA), but the role of PKA is not understood. We have demonstrated, in NC cultures, cAMP-induced transcription of the microphthalmia-associated transcription factor gene (Mitf) and the RE-1 silencing transcription factor gene (REST), both Wnt-regulated genes. In NC cultures and zebrafish, knockdown of the corepressor of Wnt-mediated transcription C-terminal binding protein 2 (CtBP2) but not CtBP1 derepressed Mitf and REST expression and enhanced melanocyte differentiation. cAMP in NC and B16 melanoma cells decreased CtBP2 protein levels, while inhibition of PKA or proteasome rescued CtBP2 degradation. Interestingly, knockdown of homeodomain-interacting protein kinase 2 (HIPK2), a CtBP stability modulator, increased CtBP2 levels, suppressed expression of Mitf, REST, and melanocyte differentiation, and increased neuronal gene expression and sympathoadrenal lineage differentiation. We conclude that cAMP/PKA via HIPK2 promotes CtBP2 degradation, leading to Mitf and REST expression. Mitf induces melanocyte specification, and REST suppresses neuron-specific gene expression and the sympathoadrenal lineage. Our studies identify a novel role for REST in NC cell differentiation and suggest cross talk between cAMP and Wnt signaling in NC lineage specification.