Alternative splicing of a carboxyl/choline esterase gene enhances the fenpropathrin tolerance of Tetranychus cinnabarinus
Alternative splicing of a carboxyl/choline esterase gene enhances the fenpropathrin tolerance of Tetranychus cinnabarinus
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DOI：
10.1111/1744-7917.13166
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发表时间：
2022-12
期刊：
影响因子：
4
通讯作者：
P. Wei;Xinying Zeng;Haonan Han;Yiqing Yang;Y. Zhang;Lin He
中科院分区：
文献类型：
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作者：
P. Wei;Xinying Zeng;Haonan Han;Yiqing Yang;Y. Zhang;Lin He
解毒作用在农业害虫抵抗农药中起着至关重要的作用，细胞色素P450、羧基/胆碱酯酶（CCE）和谷胱甘肽-S-转移酶是负责其解毒能力的主要蛋白质。CCE的活性可以通过突变上调、下调或修饰。然而，很少有研究探讨选择性剪接在改变CCE特性中的作用。我们在朱砂叶螨中鉴定了2种TcCCE 23变体：长版本（CCE 23-V1）和比CCE 23-V1短18个核苷酸的短版本（CCE 23-V2）。剪接是否影响TcCCE 23的活性仍不清楚。CCE 23 ‐V2在甲氰菊酯耐药T. cinnabarinus表明，剪接影响了CCE 23-V2对甲氰菊酯的解毒作用。当CCE 23 ‐V2的表达被敲低时，螨的死亡率显著升高（43.2% ± 3.3%）通过注射CCE 23-dsRNA（双链RNA）与在甲氰菊酯暴露下注射绿色荧光蛋白-dsRNA的对照组相比;然而，通过CCE 23-小干扰RNA下调CCE 23-V1（61.3% ± 6.3%）没有这样的效果，表明CCE 23-V2在异生物质代谢中比CCE 23-V1起更大的作用。过表达CCE 23-V2的果蝇对甲氰菊酯的耐受性（50%致死剂量[LD 50] = 19.47 μg/g）显著高于Gal 4/UAS-CCE 23-V1转基因果蝇（LD 50 = 13.11 μg/g）。分子对接分析表明，剪接打开了一扇“门”，扩大了CCE 23-V2的底物结合腔，可能增强了CCE 23-V2携带甲氰菊酯分子的能力。这些发现表明剪接可能增强TcCCE 23的解毒能力。总的来说，我们的数据提高了对CCE调节机制的多样性和复杂性的理解。
Detoxification plays a crucial role in agricultural pests to withstand pesticides, and cytochrome P450s, carboxyl/choline esterases (CCEs), and glutathione‐S‐transferases are the main proteins responsible for their detoxification ability. The activity of CCEs can be upregulated, downregulated, or modified by mutation. However, few studies have examined the role of alternative splicing in altering the properties of CCEs. We identified 2 variants of TcCCE23 in Tetranychus cinnabarinus: a long version (CCE23‐V1) and a short version that is 18 nucleotides shorter than CCE23‐V1 (CCE23‐V2). Whether splicing affects the activity of TcCCE23 remains unclear. Overexpression of CCE23‐V2 in fenpropathrin‐resistant T. cinnabarinus revealed that splicing affected the detoxification of fenpropathrin by CCE23‐V2. The mortality of mites was significantly higher when the expression of CCE23‐V2 was knocked down (43.2% ± 3.3%) via injection of CCE23‐dsRNA (double‐stranded RNA) compared with the control group injected with green fluorescent protein‐dsRNA under fenpropathrin exposure; however, the downregulation of CCE23‐V1 (61.3% ± 6.3%) by CCE23‐small interfering RNA had no such effect, indicating CCE23‐V2 plays a greater role in xenobiotic metabolism than CCE23‐V1. The tolerance of flies overexpressing CCE23‐V2 to fenpropathrin (50% lethal dose [LD50] = 19.47 μg/g) was significantly higher than that of Gal4/UAS‐CCE23‐V1 transgenic flies (LD50 = 13.11 μg/g). Molecular docking analysis showed that splicing opened a “gate” that enlarges the substrate binding cavity of CCE23‐V2, might enhance the ability of CCE23‐V2 to harbor fenpropathrin molecules. These findings suggest that splicing might enhance the detoxifying capability of TcCCE23. Generally, our data improve the understanding of the diversity and complexity of the mechanisms underlying the regulation of CCEs.