STRUCTURE AND REGULATION OF ECDYSONE-RESPONSIVE GENES

蜕皮激素反应基因的结构和调控

• 批准号: 3293568
• 负责人: Peter T CHERBAS
• 金额: $ 19.23万
• 依托单位: TRUSTEES OF INDIANA UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 1986
• 资助国家: 美国
• 起止时间: 1986-04-01 至 1989-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3293568
• 关键词: DNA; DNA footprinting; Drosophilidae; antiantibody; autoradiography; cell bank /registry; chromatin; developmental genetics; ecdysone; eukaryote; gene expression; genetic manipulation; genetic mapping; genetic promoter element; genetic recombination; genetic transcription; hormone binding protein; hormone receptor; hormone regulation /control mechanism; immunofluorescence technique; laboratory mouse; laboratory rabbit; larva; nucleic acid probes; nucleic acid sequence; radiotracer; steroid hormone

Recent studies using vertebrate systems have shown that steroid hormones, in concert with their receptor proteins, bind to DNA sequences near particular genes, modulating the activities of those gene. Ecdysone, the steroid hormone of insects, has been studied for many years, but the details of its action are as yet unknown. We propose to elucidate the mechanism of ecdysone action by identifying those DNA sequences which confer ecdysoneinducibility on particular genes (EREs) and by studying the molecules that bind to those sequences. This proposal focuses on Drosophila genes in order to facilitate the eventual genetic analysis of these regulatory pathways. The Eip28/29 and Eip40 genes are well-characterized Drosophila genes which are induced by ecdysonein several cell lines. Indirect tests suggest that they participate in the primary response of these cells to the steroid hormone. A novel situation exists at the Eip40 locus: both strands of the DNA are transcribed and both transcripts are induced during the early response to ecdysone. Remaining structural questions at the Eip40 locus will be resolved by standard procedures. These experiments should suggest whether RNA complementarity plays a regulatory role at this locus. EREs for all 3 transcription units will be mapped. Once mapped, the DNA elements will be used as probes to identify the proteins with which they interact, perhaps including the ecdysone receptor. Identification of these critical components of the response should permit the design of in vivo assays to probe their structure. Such assays might include tests of DNase and Sl hypersensitivity and in vivo footprinting. The developmental pattern of Eip gene expression in flies will be determined and will be interpreted in the light of the assays of critical structures just described. Finally, efforts will be made to uncover the functional roles of the Eip gene products. These experiments should enhance our understanding of ecdysone, of transcriptional regulation in eukaryotes, and of the relation between steroid hormones and other elements of the developmental regulatory apparatus.

使用脊椎动物系统的最新研究表明，类固醇激素， 与其受体蛋白一致，结合在附近的DNA序列 特定基因，调节这些基因的活性。 Ecdysone， 昆虫的类固醇激素已经研究了很多年，但是 其动作的细节尚不清楚。 我们建议阐明 通过识别那些DNA序列的机理 通过研究 与这些序列结合的分子。 该提议重点 果蝇基因以促进最终的遗传分析 这些监管途径。 EIP28/29和EIP40基因是特征良好的果蝇基因， 由ecdysonein诱导几种细胞系。 间接测试表明 他们参与这些细胞对类固醇的主要反应 激素。 EIP40基因座存在一种新颖的情况： DNA被转录，并在早期诱导两个转录本 对ecdysone的反应。 EIP40基因座的剩余结构性问题将由 标准程序。 这些实验应表明RNA是否 互补性在该基因座起着调节作用。 所有3个 转录单元将被映射。 映射后，DNA元素将是 用作识别与之相互作用的蛋白质的探针，也许 包括ecdysone受体。 这些关键的识别 响应的组件应允许设计体内测定 探测它们的结构。 这样的测定可能包括DNase和SL的测试 高敏性和体内足迹。 的发展模式 将确定果蝇中的EIP基因表达，并将在 刚刚描述的关键结构测定法。 最后， 将努力揭示EIP基因的功能作用 产品。 这些实验应增强我们对ecdysone的理解 真核生物中的转录调节以及 类固醇激素和发育调节的其他元素 设备。