Juvenile Hormone Receptor in Flies and Mosquitoes

描述（由申请人提供）：蜕皮酮和幼体激素（JH）是昆虫生殖和发育的主要内分泌调节剂。JH的作用机制比蜕皮酮的作用机制更不为人所知，这主要是由于我们对JH受体和JH调节基因的了解不足。黑腹果蝇（Drosophila melanogaster）耐甲氧丁烯（methopene - resistant, Met） bHLH-PAS基因参与了JH的作用，该基因可作为理解JH在黑腹果蝇和蚊子体内的分子作用的关键。这项工作是一种遗传学方法，将实现四个目标：首先，将从埃及伊蚊中分离出Met+同源基因并进行测序。Ae。埃及伊蚊是一种得到充分研究的病媒，这项工作将开始了解蚊子中的JH受体。其次，将对与Met相互作用的突变体进行增强子/抑制子筛选，以确定JH受体复合物和相关蛋白。第三，将对Met进行点突变和缺失突变，以确定Met中对该基因功能至关重要的区域。最后，通过DNA微阵列分析鉴定受JH转录控制的基因（JH靶基因）。这些基因的鉴定将有助于我们了解JH的作用，并在未来的工作中允许构建JH的报告基因测定，以促进JH信号转导的进一步分子分析。

DESCRIPTION (provided by the applicant): Ecdysterone and juvenile hormone (JH) are the primary endocrine regulators of reproduction and development in insects. The mechanism of action of JH is much more poorly understood than that of ecdysterone, due primarily to our dismal knowledge of the JH receptor and JH-regulated genes. The Methoprene-tolerant (Met) bHLH-PAS gene of Drosophila melanogaster is involved in the action of JH, and it can serve as a key to understanding the molecular action of this hormone in both D. melanogaster and mosquitoes. The proposed work is a genetic approach that will accomplish four objectives: First, the Met+ homologous gene from the aedine mosquito Aedes aegypti will be isolated and sequenced. Ae. aegypti is a well-studied disease vector, and this effort will begin an understanding of the JH receptor in mosquitoes. Second, an enhancer/suppressor screen of mutants that interact with Met will be carried out to define the JH receptor complex and related proteins. Third, both point and deletion mutagenesis of Met will be carried out to define the region(s) of MET critical for the functions of this gene. Finally, genes that are transcriptionally controlled by JH (JH target genes) will be identified by DNA micro-array analysis. Identification of these genes will help us understand JH roles and in future work will permit construction of a reporter gene assay for JH to facilitate further molecular analysis of JH signal transduction. This work will have significance for understanding the JH receptor in a genetically well-defined insect. It will lead to a better understanding of the action and insect resistance to the insecticide methoprene, which is used to control mosquito vectors of disease. In addition, a better understanding of Met as a bHLH-PAS gene will further our knowledge of this family of transcriptional regulators, including homologous genes found in humans.