E63-1 AND STEROID/CALCIUM-REGULATED CROSSTALK

E63-1 和类固醇/钙调节串扰

• 批准号: 2771047
• 负责人: ANDREW J ANDRES
• 金额: $ 14.95万
• 依托单位: NORTHWESTERN UNIVERSITY AT CHICAGO
• 依托单位国家: 美国
• 财政年份: 1997
• 资助国家: 美国
• 起止时间: 1997-09-15 至 2001-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2771047
• 关键词: Drosophilidae; binding proteins; biological signal transduction; calcium binding protein; calcium flux; calcium ion; calmodulin dependent protein kinase; developmental genetics; ecdysone; gene expression; genetic library; genetic manipulation; genetically modified animals; hormone regulation /control mechanism; larva; micromanipulator; molecular cloning; physiology; polymerase chain reaction; protein structure function; salivary glands; secretory immune system

Steroid hormones are small regulatory molecules that profoundly influence basic processes of growth and homeostasis in developing organisms. Presumably, this occurs when the hormone induces a qualitative change in the pattern of gene expression. Thus, identifying and characterizing the gene targets for steroid action, and how they are translated into biological changes in the target tissue, is a primary goal of this research. Specifically, this proposal is a logical extension of the unexpected discovery made during the PI's post-doctoral studies on tissue-specific (salivary gland) steroid-regulated genes in the fruit fly Drosophila melanogaster. Briefly, it was shown that E63-1, a primary (early) target for the Drosophila steroid ecdysone, encodes a high-affinity Ca2+-binding protein. This discovery was novel and exciting because most of the previously characterized early targets were shown to encode transcription factors. This finding provides the first link between steroid-regulated and Ca2+-regulated signaling pathways in Drosophila. Thus, one is afforded a unique opportunity to investigate this possible "crosstalk" in a genetically tractable organism in the context of an already well characterized molecular hierarchy. The overall hypothesis being addressed is that E63-1 functions to coordinate a steroid- and calcium-mediated regulatory pathway in the salivary gland. The goals are to dissect this pathway at the molecular level, investigate the possibility that E64-1 encodes a signal transduction molecule, and apply this information to other systems. The specific aims are as follows: 1) Genetically manipulate the levels of E63-1 protein and investigate these effects on the physiological state of the tissue (secretory activity, gene expression manifested as puffs, intracellular Ca2+ changes). 2) Further define the mechanism by initiating a molecular screen for downstream acceptor proteins of E63-1.

类固醇激素是一种小调节分子，对 影响发育中生长和体内平衡的基本过程 有机体。 据推测，当激素诱导 基因表达模式发生质的变化。 因此， 识别和表征类固醇作用的基因靶标， 以及它们如何转化为目标的生物变化 组织，是这项研究的主要目标。 具体而言，本提案 是对期间意外发现的逻辑延伸 PI组织特异性（唾液腺）博士后研究 果蝇果蝇中的类固醇调节基因。 简而言之，研究表明 E63-1 是 果蝇类固醇蜕皮激素，编码高亲和力 Ca2+ 结合 蛋白质。 这个发现是新颖且令人兴奋的，因为大多数 先前表征的早期目标被证明可以编码 转录因子。 这一发现提供了第一个联系 类固醇调节和 Ca2+ 调节的信号通路 果蝇。 因此，人们获得了一个独特的机会 研究遗传易处理的这种可能的“串扰” 在已经充分表征的分子背景下的有机体 等级制度。 所讨论的总体假设是 E63-1 的功能是 协调类固醇和钙介导的调节途径 唾液腺。 目标是剖析这条路径 分子水平，研究E64-1编码a的可能性 信号转导分子，并将此信息应用于其他 系统。 具体目标如下： 1) 基因操纵E63-1蛋白的水平和 研究这些对组织生理状态的影响 （分泌活性，基因表达表现为泡芙， 细胞内 Ca2+ 变化）。 2）通过启动分子筛选进一步明确机制 用于 E63-1 的下游受体蛋白。