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    • A Novel GSTe2 Involved in Metamorphosis by Regulating 20E Signal Pathway in Tribolium Castaneum.

    KumChol Kim,XiaoWen Song,RunNan Yu,YongLei Zhang,Han Gao,SuiSui Wang,Bin Li

    Archives Of Insect Biochemistry And Physiology（2023）SCI 4区

    Department of Life-Science | Jiangsu Key Laboratory for Biodiversity and Biotechnology

    Cited 4|Views9
    Abstract
    Insect-specific epsilon glutathion S-transferases (GSTs) are a class of multifunctional GST superfamily, which play important roles in detoxification of xenobiotic substances. Most research on GSTs has focused on insecticide detoxification and resistance, with little research on other physiological functions. Here, we identified and cloned the novel GSTe2 from Tribolium castaneum (TcGSTe2). Recombinant TcGSTe2 protein was successfully overexpressed in Escherichia coli and purified with affinity purification, which had high ability to catalyze the conjugation of reduced glutathione with 1-chloro-2,4-dinitrobenzene (CDNB). The expression level of TcGSTe2 was significantly decreased after exposure with four insecticides, phoxim, λ-cyhalothrin, dichlorvos, and carbofuran, in larval stage. Interestingly, RNA interference knockdown of TcGSTe2 caused metamorphosis deficiency in larval and pupal stages by inhibiting the 20E signal pathway. Furthermore, exogenous 20E injection partially rescued this metamorphosis deficiency and also increased the expression levels of 20E downstream response genes. This study illustrated TcGSTe2 plays an important role at metamorphosis beside the insecticide detoxification and resistance in T. castaneum.
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    20E pathway,ecdysteroid,glutathione S-transferase,metamorphosis,recombinant protein,Tribolium castaneum
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    【要点】：本研究发现了一种新型GSTe2（TcGSTe2）在调节20E信号途径中参与调节赤拟谷盗（Tribolium castaneum）的变态过程，揭示了GSTs在昆虫生理功能中的新作用。
    

    【方法】：通过基因克隆和重组蛋白表达技术，从赤拟谷盗中鉴定并克隆了TcGSTe2，并在大肠杆菌中成功过表达了该蛋白。
    

    【实验】：通过亲和纯化得到TcGSTe2蛋白，发现其具有催化还原型谷胱甘肽与1-氯-2,4-二硝基苯（CDNB）结合的高能力。实验使用四种杀虫剂（phoxim、λ-cyhalothrin、dichlorvos和carbofuran）处理幼虫，发现TcGSTe2表达水平显著下降。通过RNA干扰技术敲除TcGSTe2，导致幼虫和蛹期变态缺陷，且外源性20E注射部分恢复了这种变态缺陷，并提高了20E下游响应基因的表达水平。实验使用的数据集为赤拟谷盗幼虫和蛹期的TcGSTe2表达数据。