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首页> 外文期刊>Bioscience Reports >Transcription of the chicken Grin1 gene is regulated by the activity of SP3 and NRSF in undifferentiated cells and neurons
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Transcription of the chicken Grin1 gene is regulated by the activity of SP3 and NRSF in undifferentiated cells and neurons

机译：鸡Grin1基因的转录受未分化细胞和神经元中SP3和NRSF活性的调节

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The NMDA (N-methyl-D-aspartate) receptors are important in the regulation of neuronal development, synaptic plasticity, learning and memory, and are involved in several brain pathologies. The NR1 subunit is essential for the assembly of functional receptors, as it forms the calcium-permeable ion channel and contains the obligatory co-agonist binding site. Previous studies have shown that NR1 gene (Grin1) expression is up-regulated during neuronal differentiation and its expression is widespread in the central nervous system. We have previously cloned the chicken Grin1 gene and 1.9?kb of the 5′-regulatory region. In the present study, we analysed the molecular mechanisms that regulate chicken Grin1 gene transcription in undifferentiated cells and neurons. By functional analysis of chicken Grin1–luciferase gene 5′-regulatory region constructs, we demonstrate that the basal promoter is delimited within 210?bp upstream from the main transcription initiation site. DNA–protein binding and functional assays revealed that the 5′-UTR (untranslated region) has one consensus NRSE (neuron-restrictive silencing element) that binds NRSF (neuron-restrictive silencing factor), and one SP (stimulating protein transcription factor) element that binds SP3, both repressing Grin1 gene transcription in undifferentiated P19 cells (embryonic terato-carcinoma cells) and PC12 cells (phaeochromocytoma cells). The promoter region lacks a consensus TATA box, but contains one GSG/SP (GSG-like box near a SP-consensus site) that binds SP3 and up-regulates gene transcription in embryonic chicken cortical neurons. Taken together, these results demonstrate a dual role of SP3 in regulating the expression of the Grin1 gene, by repressing transcription in the 5′-UTR in undifferentiated cells as well as acting as a transcription factor, increasing Grin1 gene transcription in neurons.
机译：NMDA（N-甲基-D-天冬氨酸）受体在神经元发育，突触可塑性，学习和记忆的调节中很重要，并涉及几种脑部疾病。 NR1亚基对于功能受体的组装是必不可少的，因为它形成了钙可渗透的离子通道，并包含强制性的激动剂结合位点。先前的研究表明，NR1基因（Grin1）的表达在神经元分化过程中被上调，并且其表达在中枢神经系统中广泛分布。我们以前已经克隆了鸡Grin1基因和5'-调控区的1.9？kb。在本研究中，我们分析了调节未分化细胞和神经元中鸡Grin1基因转录的分子机制。通过对鸡Grin1-荧光素酶基因5'-调节区构建体的功能分析，我们证明了基础启动子在主要转录起始位点上游210？bp内被定界。 DNA-蛋白质结合和功能测定表明，5'-UTR（非翻译区）具有一个结合了NRSF（神经限制沉默因子）的共有NRSE（神经限制沉默元件）和一个SP（刺激蛋白质转录因子）元件结合SP3，可抑制未分化的P19细胞（胚胎性畸胎癌细胞）和PC12细胞（嗜铬细胞瘤细胞）中的Grin1基因转录。启动子区域没有一个共有的TATA框，但包含一个GSG / SP（一个位于SP共有位点附近的GSG样框），它结合了SP3并上调了胚胎鸡皮质神经元的基因转录。综上所述，这些结果证明了SP3通过抑制未分化细胞中5'-UTR中的转录并作为转录因子，增加神经元中Grin1基因的转录，从而调节Grin1基因的表达。

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