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Juvenile hormone regulation of an insect gene: a specific transcription factor and a DNA response element.

机译:昆虫基因的少年激素调节:特定的转录因子和DNA反应元件。

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摘要

We have used locust fat body nuclear protein extracts and upstream DNA of the juvenile hormone (JH)-inducible locust gene, jhp21, to examine the regulation of specific transcription by JH. Promoter activity was assayed with G-free cassette reporter constructs. Nuclear extracts from adult female fat body, previously exposed to JH or an analog, actively transcribe from the jhp21 promoter and a control adenovirus major late (AdML) promoter, whereas extracts from JH-deprived female fat body, or other tissues, transcribe strongly from the AdML promoter but weakly or not at all from the jhp21 promoter. Transcription is enhanced by sequences between -140 and -211 nt from the jhp21 transcription start point (tsp), which include a CAAT box, and also by sequences between -1056 and -1200. A 15-nt partially palindromic sequence element found at -1152, resembling known hormone response elements, was shown to stimulate transcription when restored to truncated jhp21 DNA. Two very similar sequences occur further upstream. In electrophoretic mobility shift assays (EMSA), the same sequence element was shown to specifically bind a protein that was present in nuclear extracts from JH-exposed, but not from JH-deprived, fat body. Several lines of evidence suggest that the DNA element may be a JH response element (JHRE). The JH-induced protein that binds to it appears to be a transcription factor that activates the initiation of JH target gene (jhp21) transcription, and could be a JH receptor.
机译:我们已经使用了蝗虫脂肪核蛋白提取物和可诱导少年激素(JH)的蝗虫基因jhp21的上游DNA,来检查JH特异性转录的调控。用无G的盒报道基因构建物测定启动子活性。先前暴露于JH或类似物的成年雌性脂肪体的核提取物从jhp21启动子和对照腺病毒主要晚期(AdML)启动子中主动转录,而JH缺乏的雌性脂肪体或其他组织的提取物从AdML启动子,但jhp21启动子几乎没有或根本没有。从jhp21转录起点(tsp)开始的-140和-211 nt之间的序列(包括CAAT框)以及-1056和-1200之间的序列增强了转录。发现在-1152处发现的15 nt部分回文序列元件类似于已知的激素反应元件,当恢复为截短的jhp21 DNA时,可刺激转录。在上游有两个非常相似的序列。在电泳迁移率变动分析(EMSA)中,显示了相同的序列元件特异性结合存在于暴露于JH的脂肪提取物中但不来自暴露于JH的脂肪体的核提取物中的蛋白质。几条证据表明,DNA元件可能是JH反应元件(JHRE)。 JH诱导的与其结合的蛋白似乎是激活JH目标基因(jhp21)转录起始的转录因子,并且可能是JH受体。

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