REST-VP16 activates multiple neuronal differentiation genes in human NT2 cells
REST-VP16 activates multiple neuronal differentiation genes in human NT2 cells
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DOI：
10.1093/nar/28.17.3403
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发表时间：
2000-09-01
影响因子：
14.9
通讯作者：
Majumder, S
中科院分区：
文献类型：
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作者：
Immaneni, A;Lawinger, P;Majumder, S
Re1沉默转录因子(REST)/神经元限制性沉默因子(NRSF)通过与非神经细胞中存在的特定的共同DNA序列结合来抑制非神经细胞中一系列神经元分化基因的转录。然而，REST/NRSF-/-小鼠表明，仅缺乏REST/NRSF依赖的抑制是不足以表达这些神经元分化基因的，还需要其他启动子/增强子特异性激活剂的存在。在这里，我们描述了重组转录因子REST-VP16的构建，通过用病毒激活因子VP16的激活结构域取代REST/NRSF的抑制域，在瞬时转染实验中，发现REST-VP16通过Re1结合位点/神经元限制性增强子元件(Re1/NRSE)起作用，在各种哺乳动物细胞中激活质粒编码的神经元启动子，并激活细胞REST/NRSF靶基因，即使在缺乏激活这些基因所需的因子的情况下也是如此。通过腺病毒载体在NT2细胞中高效表达REST-VP16被发现能引起多个神经元基因的激活，这些基因是神经元分化的特征标志。因此，REST-VP16可以作为一种独特的工具来研究神经元分化途径和由于这一过程的放松而引起的神经元疾病。
The RE1-silencing transcription factor (REST)/neuron-restrictive silencer factor (NRSF) can repress transcription of a battery of neuronal differentiation genes in non-neuronal cells by binding to a specific consensus DNA sequence present in their regulatory regions. However, REST/NRSF-/- mice suggest that the absence of REST/NRSF-dependent repression alone is not sufficient for the expression of these neuronal differentiation genes and that the presence of other promoter/enhancer-specific activators is required. Here we describe the construction of a recombinant transcription factor, REST-VP16, by replacing repressor domains of REST/NRSF with the activation domain of a viral activator VP16, In transient transfection experiments, REST-VP16 was found to operate through RE1 binding site/neuron-restrictive enhancer element (RE1/NRSE), activate plasmid-encoded neuronal promoters in various mammalian cell types and activate cellular REST/ NRSF target genes, even in the absence of factors that are otherwise required to activate such genes. Efficient expression of REST-VP16 through adenoviral vectors in NT2 cells, which resemble human committed neuronal progenitor cells, was found to cause activation of multiple neuronal genes that are characteristic markers for neuronal differentiation. Thus, REST-VP16 could be used as a unique tool to study neuronal differentiation pathways and neuronal diseases that arise due to the deregulation of this process.