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首页> 外文期刊>Life sciences >Regulation of cholinergic gene expression by the neuron restrictive silencer factor/repressor element-1 silencing transcription factor.
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Regulation of cholinergic gene expression by the neuron restrictive silencer factor/repressor element-1 silencing transcription factor.

机译：神经元限制性沉默子因子/阻遏物1沉默转录因子对胆碱能基因表达的调节。

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摘要

The role of protein kinase A in regulating transcription of the cholinergic gene locus, which contains both the vesicular acetylcholine transporter gene and the choline acetyltransferase gene, was investigated in PC12 cells and a protein kinase A deficient PC12 mutant, A126.1B2 in which transcription of the locus is reduced. The site of action of protein kinase A was localized to a neuron restrictive silencer element/repressor element-1 (NRSE/RE-1) within the upstream region of the cholinergic gene locus. The neuron restrictive silencer factor/repressor element-1 silencing transcription factor (NRSF/REST), the transcription factor which binds to NRSE/RE-1, was expressed at similar levels in both PC12 and A126.1B2. Although nuclear extracts containing NRSF/REST from A126.1B2 exhibited binding to NRSE/RE-1, nuclear extracts from PC12 cells did not. The NRSF/REST isoform repressor element-1 silencing transcription factor-4 (REST4) was found to be expressed in PC12 cells, but not in the protein kinase A deficient PC12 cell line. REST4 inhibited the binding of NRSF/REST to NRSE/RE-1 as determined by gel mobility shift assays. Co-immunoprecipitation was used to demonstrate interaction between NRSF/REST and REST4. Expression of recombinant REST4 in the protein kinase A deficient PC12 cell line was sufficient to transcriptionally activate the cholinergic gene locus. Thus in PC12 cells protein kinase A promotes the production of REST4, which in turn de-represses of the cholinergic gene locus by inactivating the transcription repressor NRSF/REST.
机译：在PC12细胞和蛋白激酶A缺陷型PC12突变体A126.1B2中，研究了蛋白激酶A在调节胆碱能基因位点的转录中的作用，该位点既包含水泡乙酰胆碱转运蛋白基因又包含胆碱乙酰基转移酶基因。轨迹减少。蛋白激酶A的作用位点位于胆碱能基因位点上游区域内的神经元限制性沉默子元件/阻遏物1（NRSE / RE-1）。与NRSE / RE-1结合的神经元限制性沉默沉默因子/阻遏物1沉默转录因子（NRSF / REST）在PC12和A126.1B2中的表达水平相似。尽管含有来自A126.1B2的NRSF / REST的核提取物表现出与NRSE / RE-1的结合，但来自PC12细胞的核提取物却没有。发现NRSF / REST亚型阻遏元件1沉默转录因子4（REST4）在PC12细胞中表达，但在蛋白激酶A缺陷型PC12细胞系中不表达。 REST4抑制了NRSF / REST与NRSE / RE-1的结合，这是通过凝胶迁移率迁移分析确定的。共免疫沉淀被用来证明NRSF / REST和REST4之间的相互作用。重组REST4在蛋白激酶A缺失的PC12细胞系中的表达足以转录激活胆碱能基因位点。因此，在PC12细胞中，蛋白激酶A促进REST4的产生，而REST4则通过使转录阻遏物NRSF / REST失活而降低了胆碱能基因位点的抑制。

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