Juvenile Hormone Receptor in Flies and Mosquitoes

描述(申请人提供)：蜕皮激素和保幼激素(JH)是昆虫繁殖和发育的主要内分泌调节剂。JH的作用机制远比蜕皮酮知之甚少，这主要是因为我们对JH受体和JH调节基因的了解很少。果蝇耐甲基丁二烯(Met)bHLH-PAS基因参与了JH的作用，可作为了解该激素在黑腹果蝇和蚊子中的分子作用的关键。这项拟议的工作是一种遗传学方法，将实现四个目标：首先，将分离埃及伊蚊Met+同源基因并进行序列测定。天哪。埃及伊蚊是一种经过充分研究的疾病媒介，这一努力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，以定义JH受体复合体和相关蛋白。第三，Met基因的点突变和缺失突变都将被用来确定Met基因的关键区域(S)。最后，将通过DNA微阵列分析来鉴定受JH(JH靶基因)转录控制的基因。这些基因的鉴定将有助于我们了解JH的作用，并在未来的工作中将有助于建立JH的报告基因分析，以便于进一步的JH信号转导的分子分析。 这项工作将对理解基因定义明确的昆虫中的JH受体具有重要意义。这将有助于更好地了解杀虫剂甲氧丁二胺的作用和对昆虫的抗药性，该杀虫剂用于控制蚊子传播疾病的媒介。此外，更好地理解Met作为bHLH-PAS基因将促进我们对这一转录调控家族的了解，包括在人类中发现的同源基因。

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.