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首页> 外文期刊>Molecular and Cellular Biology >Distinct cis-acting elements mediate clock, light, and developmental regulation of the Neurospora crassa eas (ccg-2) gene.
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Distinct cis-acting elements mediate clock, light, and developmental regulation of the Neurospora crassa eas (ccg-2) gene.

机译：不同的顺式作用元件介导了神经孢霉（ccg-2）基因的时钟，光和发育调控。

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摘要

The Neurospora crassa eas (ccg-2) gene, which encodes a fungal hydrophobin, is transcriptionally regulated by the circadian clock. In addition, eas (ccg-2) is positively regulated by light and transcripts accumulate during asexual development. To sort out the basis of this complex regulation, deletion analyses of the eas (ccg-2) promoter were carried out to localize the cis-acting elements mediating clock, light, and developmental control. The primary sequence determinants of a positive activating clock element (ACE) were found to reside in a 45-bp region, just upstream from the TATA box. Using a novel unregulated promoter/reporter system developed for this study, we show that a 68-bp sequence encompassing the ACE is sufficient to confer clock regulation on the eas (ccg-2) gene. Electrophoretic mobility shift assays using the ACE reveal factors present in N. crassa protein extracts that recognize and bind specifically to DNA containing this element. Separate regions of the eas (ccg-2) promoter involved in light induction and developmental control are identified and shown not to be required for clock-regulated expression of eas (ccg-2). The distinct nature of the ACE validates its use as a tool for the identification of upstream regulatory factors involved in clock control of gene expression.
机译：昼夜节律在神经真菌孢子（ccg-2）基因上编码真菌疏水蛋白。另外，eas（ccg-2）受光正调控，转录本在无性发育过程中积累。为了梳理这种复杂调控的基础，对eas（ccg-2）启动子进行了缺失分析，以定位介导时钟，光照和发育控制的顺式作用元件。发现正激活时钟元件（ACE）的主要序列决定簇位于TATA盒上游45 bp区域。使用为该研究开发的新型不受调控的启动子/报告系统，我们表明包含ACE的68 bp序列足以赋予eas（ccg-2）基因时钟调节。使用ACE的电泳迁移率变化分析揭示了crassa猪笼草蛋白质提取物中存在的因子，这些因子可识别并特异性结合包含该元素的DNA。鉴定了与光诱导和发育控制有关的eas（ccg-2）启动子的单独区域，并显示它不是时钟调控的eas（ccg-2）表达所必需的。 ACE的独特性质验证了其作为鉴定与基因表达的时钟控制有关的上游调节因子的工具的用途。

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