INTERACTION OF PROTEINS INVOLVED IN ECDYSONE AND JUVENILE HORMONE SIGNAL TRANSDUCTION
INTERACTION OF PROTEINS INVOLVED IN ECDYSONE AND JUVENILE HORMONE SIGNAL TRANSDUCTION
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DOI：
10.1002/arch.20281
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发表时间：
2009-02-01
影响因子：
2.2
通讯作者：
Palli, Subba Reddy
中科院分区：
文献类型：
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作者：
Bitra, Kavita;Palli, Subba Reddy
蜕皮激素和保幼激素（JH）调节各种发育，生理，行为和代谢过程。蜕皮激素通过两种核受体的异二聚体复合物发挥作用，即蜕皮激素受体（EcR）和超气门（USP）。在果蝇耐甲氧普烯（Met）突变体中鉴定的85 kDa蛋白与JH III高亲和力结合，并且突变体果蝇对保幼激素类似物（JHA）甲氧普烯具有抗性。为了研究EcR、USP和Met之间的分子相互作用，进行了使用酵母双杂交系统的报告基因测定。正如预期，在存在稳定蜕皮甾类类似物、RG-102240或甾类化合物、鼠甾酮A和泊那甾酮A的情况下，与B42活化结构域融合的EcR和与莱克萨DNA结合结构域融合的USP相互作用，并支持报告基因的诱导。USP：USP同源二聚体支持在不存在配体的情况下报告基因的表达，并且在添加JHA、甲氧普烯后报告基因活性没有显著增加。同样，Met：Met同源二聚体以及Met：EcR和Met：USP异源二聚体在不存在配体的情况下诱导报告活性，并且添加蜕皮甾类或JH类似物并不增加由Met蛋白的同源二聚体或异源二聚体调节的报告活性。昆虫细胞中的双杂交试验和体外下拉试验证实了Met与EcR和USP的相互作用。这些数据表明，参与蜕皮激素（EcR和USP）和保幼激素（Met）信号转导的蛋白质相互作用，介导这两种重要激素之间的串扰。(C)2008年威利期刊公司
Ecdysteriods and juvenile hormones (JH) regulate a variety of developmental, physiological, behavioral, and metabolic processes. Ecdysteriods function through a heterodimeric complex of two nuclear receptors, ecdysone receptor (EcR) and ultraspiracle (USP). An 85 kDa protein identified in Drosophila melanogaster methoprene-tolerant (Met) mutant binds to JH III with high affinity, and the mutant flies are resistant to juvenile hormone analog (JHA), methoprene. Reporter assays using the yeast two-hybrid system were performed in order to study the molecular interactions between EcR, USP and Met. As expected, EcR fused to the B42 activation domain and USP fused to the LexA DNA binding domain interacted with each other and supported induction of the reporter gene in the presence of stable ecdysteriod analong, RG-102240 or steriods, muristerone A and ponasterone A. The USP:USP homodimers supported expression of the reporter gene in the absence of ligand, and there was no significant increase in the reporter activity after addition of a JHA, methoprene. Similarly, Met: Met homodimers as well as Met:EcR and Met: USP heterodimers induced reporter activity in the absence of ligand and addition of ecdysteroid or JH analogs did not increase the reporter activity regulated by either homodimers or heterodimers of Met protein. Two-hybrid assays in insect cells and in vitro pull-down assays confirmed the interaction of Met with EcR and USP. These data suggest that the proteins that are involved in signal transduction of ecdysteroids (EcR and USP) and juvenile hormones (Met) interact to mediate cross-talk between these two important hormones. (C) 2008 Wiley Periodical Inc.