Expression, subcellular localization and protein-protein interaction of four isoforms of EcR/USP in the common cutworm  

作　　者：Li-Xia Huang Yan-Jun Gong Jun Gu Bao-Juan Zeng Li-Hua Huang Qi-Li Feng 

机构地区：[1]Laboratory of Molecular and Developmental Entomology, Guangdong Provincial Key Laboratory of Biotechnology for Plant Development,School of Life Sciences, South China Normal University, Guangzhou, China

出　　处：《Insect Science》2015年第1期95-105,共11页昆虫科学（英文版）

基　　金：Acknowledgments The research was supported by the grants from National Natural Science Foundation of China （Grant No. 31172154） and the National Basic Research Program of China （973 Program, No. 2012CB114101）.

摘　　要：Ecdysone receptor （EcR） and ultraspiracle （USP） form heterodimers to mediate ecdysteroid signaling during molting and metamorphosis. Various EcR/USP heterodimers have been reported. However, it is unclear what kind of EcR/USP combination is adopted by lepidopteran insects during the larval-pupal metamorphosis and whether the EcR/USP heterodimer varies among different tissues. To address these questions, two isoforms of each EcR and USP were cloned from the common cutworm, their messenger RNA expression patterns were examined by real-time quantitative polymerase chain reaction in different tissues during the larval-pupal metamorphosis and in the midgut in response to hormonal induction. Furthermore, their subcellular localization and protein-protein interaction were explored by transient expression and far-western blotting, respectively. All the four genes were significantly up-regulated in prepuae and/or pupae. The expression profiles of EcRB1 and USP1 were nearly identical to each other in the epidermis, fat body and midgut, and a similar situation also applied to EcRA and USP2. The three genes responded to 20-hydroxyecdysone （20E） induction except for USP2, and USP1 could be up-regulated by both 20E and juvenile hormone. The four proteins mainly localized in the nucleus and the nuclear localization was promoted by 20E. The protein-protein interaction between each EcR and USP was found in vitro. These results suggest that two types of EcR/USP heterodimer （EeRA/USP2 and EcRB 1/USP1） may exist simultaneously in the common cutworm, and the latter should play more important roles during the larval-pupal metamorphosis. In addition, the types of EcR/USP heterodimer do not vary in the tissues which undergo histolysis and regeneration during metamorphosis.蜕皮激素受体(EcR ) 和 ultraspiracle (USP ) 形成 heterodimers 调停 ecdysteroid 在 molting 和变形期间发信号。各种各样的 EcR/USP heterodimers 被报导了。然而，联合什么样的 EcR/USP 被鳞翅类的昆虫在幼虫蛹的变形期间采用并且 EcR/USP heterodimer 是否在不同纸巾之中变化，是不清楚的。探讨这些问题，每 EcR 和 USP 的二 isoforms 从普通糖蛾被克隆，他们的送信人 RNA 表示模式被即时量的聚合酶链反应响应神经质的正式就职在幼虫蛹的变形期间并且在 midgut 在不同纸巾检验。而且，他们的 subcellular 本地化和蛋白质蛋白质相互作用被短暂表示并且分别地的远西方的弄污探索。所有四基因在 prepuae 或蛹是显著地起来调整的。EcRB1 和 USP1 的表示侧面与在外皮，胖身体和 midgut 的对方将近相同，并且一种类似的状况也适用于 EcRA 和 USP2。三基因除了 USP2，和 USP1 回答了正式就职到 20-hydroxyecdysone (20E ) 能由 20E 和少年荷尔蒙起来调整。在原子核和原子本地化主要局部性的四蛋白质被 20E 支持。在每 EcR 和 USP 之间的蛋白质蛋白质相互作用在 vitro 被发现。这些结果建议 EcR/USP heterodimer (EcRA/USP2 和 EcRB1/USP1 ) 的二种类型可以在普通糖蛾同时存在，并且后者应该在幼虫蛹的变形期间起更重要的作用。另外， EcR/USP heterodimer 的类型不在在变形期间经历 histolysis 和新生的纸巾变化。

关 键 词：ecdysone receptor protein interaction subcellular localization ULTRASPIRACLE 

分 类 号：Q51[生物学—生物化学] Q75