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保幼激素反应元件及其结合蛋白的鉴定与表征

Identification and characterization of a juvenile hormone response element and its binding proteins.

作者信息

Li Yiping, Zhang Zhaolin, Robinson Gene E, Palli Subba R

机构信息

Department of Entomology, University of Kentucky, Lexington, KY 40546, USA.

出版信息

J Biol Chem. 2007 Dec 28;282(52):37605-17. doi: 10.1074/jbc.M704595200. Epub 2007 Oct 22.

Abstract

Juvenile hormones (JH) regulate a wide variety of developmental and physiological processes in insects. Comparison of microarray data on JH-induced genes in the fruit fly, Drosophila melanogaster, L57 cells and in the honey bee, Apis mellifera, identified 16 genes that are induced in both species. Analysis of promoter regions of these 16 D. melanogaster genes identified DmJHRE1 (D. melanogaster JH response element 1). In L57 cells, the reporter gene regulated by DmJHRE1 was induced by JH III. Two proteins (FKBP39 and Chd64) that bind to DmJHRE1 were identified. FKBP39 and Chd64 double-stranded RNA inhibited JH III induction of a reporter gene regulated by DmJHRE1. FKBP39 and Chd64 proteins expressed in yeast bound to DmJHRE1. Two-hybrid and pull-down assays showed that these two proteins interact with each other as well as with ecdysone receptor, ultraspiracle, and methoprene-tolerant protein. Developmental expression profiles and JH induction of mRNA for FKBP39 and Chd64 proteins and their interaction with proteins known to be involved in both JH (methoprene-tolerant protein) and ecdysteroid action (ecdysone receptor and ultraspiracle) suggest that these proteins probably play important roles in cross-talk between JH and ecdysteroids.

摘要

保幼激素(JH)调节昆虫体内多种发育和生理过程。通过比较果蝇(Drosophila melanogaster)L57细胞和蜜蜂(Apis mellifera)中保幼激素诱导基因的微阵列数据,鉴定出16个在这两个物种中均被诱导的基因。对这16个黑腹果蝇基因的启动子区域进行分析,鉴定出DmJHRE1(黑腹果蝇保幼激素反应元件1)。在L57细胞中,由DmJHRE1调控的报告基因被保幼激素III诱导。鉴定出两种与DmJHRE1结合的蛋白质(FKBP39和Chd64)。FKBP39和Chd64双链RNA抑制了由DmJHRE1调控的报告基因的保幼激素III诱导作用。在酵母中表达的FKBP39和Chd64蛋白与DmJHRE1结合。双杂交和下拉实验表明,这两种蛋白质相互作用,并且与蜕皮激素受体、超气门蛋白和抗蚊蝇醚蛋白相互作用。FKBP39和Chd64蛋白的发育表达谱、mRNA的保幼激素诱导作用以及它们与已知参与保幼激素(抗蚊蝇醚蛋白)和蜕皮甾体作用(蜕皮激素受体和超气门蛋白)的蛋白质的相互作用表明,这些蛋白质可能在保幼激素和蜕皮甾体之间的相互作用中发挥重要作用。

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