Dynamic regulation of neuronal NO synthase transcription by calcium influx through a CREB family transcription factor-dependent mechanism
Dynamic regulation of neuronal NO synthase transcription by calcium influx through a CREB family transcription factor-dependent mechanism
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DOI：
10.1073/pnas.97.15.8617
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发表时间：
2000-07-18
影响因子：
11.1
通讯作者：
Dawson, TM
中科院分区：
文献类型：
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作者：
Sasaki, M;Gonzalez-Zulueta, M;Dawson, TM
神经元型一氧化氮合酶（nNOS）在多种生理和病理刺激下受到动态调节。虽然nNOS的动态调节已经建立，但这些不同刺激调节nNOS表达的分子机制尚未确定。我们描述的实验表明，通过电压敏感的Ca 2+通道的Ca 2+进入调节nNOS的表达，通过在皮层神经元中的交替启动子的使用和nNOS外显子2包含的调控序列，响应Ca 2+。nNOS外显子2启动子的缺失和突变分析揭示了两个关键的cAMP/Ca 2+反应元件（克雷斯），它们紧邻转录起始位点的上游。CREB与nNOS基因内的克雷斯结合。nNOS克雷斯的突变以及CREB功能的阻断导致nNOS转录的显著损失。这些结果表明，nNOS是一个Ca 2+调节基因，通过CREB对克雷斯的nNOS外显子2启动子内的相互作用，这些相互作用可能是集中参与调节nNOS在神经元损伤和活动依赖性可塑性。
Neuronal nitric oxide (NO) synthase (nNOS) is dynamically regulated in response to a variety of physiologic and pathologic stimuli. Although the dynamic regulation of nNOS is well established, the molecular mechanisms by which such diverse stimuli regulate nNOS expression have not yet been identified. We describe experiments demonstrating that Ca2+ entry through voltage-sensitive Ca2+ channels regulates nNOS expression through alternate promoter usage in cortical neurons and that nNOS exon 2 contains the regulatory sequences that respond to Ca2+. Deletion and mutational analysis of the nNOS exon 2 promoter reveals two critical cAMP/Ca2+ response elements (CREs) that are immediately upstream of the transcription start site. CREB binds to the CREs within the nNOS gene. Mutation of the nNOS CREs as well as blockade of CREB function results in a dramatic loss of nNOS transcription. These findings suggest that nNOS is a Ca2+-regulated gene through the interactions of CREB on the CREs within the nNOS exon 2 promoter and that these interactions are likely to be centrally involved in the regulation of nNOS in response to neuronal injury and activity-dependent plasticity.