Kim KumChol, Song XiaoWen, Yu RunNan, Zhang YongLei, Gao Han, Wang SuiSui, Li Bin
Jiangsu Key Laboratory for Biodiversity and Biotechnology, College of Life Sciences, Nanjing Normal University, Nanjing, China.
Department of Life-Science, University of Science, Pyongyang, Democratic People's Republic of Korea.
Arch Insect Biochem Physiol. 2023 Mar;112(3):e21989. doi: 10.1002/arch.21989. Epub 2023 Jan 1.
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.
昆虫特异性ε-谷胱甘肽S-转移酶（GSTs）是多功能GST超家族的一类，在异源生物物质的解毒中发挥重要作用。大多数关于GSTs的研究都集中在杀虫剂解毒和抗性方面，而对其他生理功能的研究较少。在此，我们从赤拟谷盗中鉴定并克隆了新型GSTe2（TcGSTe2）。重组TcGSTe2蛋白在大肠杆菌中成功过表达，并通过亲和纯化进行了纯化，其具有高催化还原型谷胱甘肽与1-氯-2,4-二硝基苯（CDNB）结合的能力。在幼虫阶段用四种杀虫剂（辛硫磷、高效氯氟氰菊酯、敌敌畏和呋喃丹）处理后，TcGSTe2的表达水平显著降低。有趣的是，通过RNA干扰敲低TcGSTe2会抑制20E信号通路，从而导致幼虫和蛹期的变态缺陷。此外，外源注射20E可部分挽救这种变态缺陷，并提高20E下游反应基因的表达水平。本研究表明，TcGSTe2在赤拟谷盗的变态过程中除了杀虫剂解毒和抗性外还发挥重要作用。
