The Metarhizium anisopliae Toxin, Destruxin A, Interacts with the SEC23A and TEME214 Proteins of Bombyx mori

文献类型： 外文期刊

第一作者： Yin, Fei

作者： Yin, Fei;Xiao, Miaomiao;Hu, Qiongbo;Yin, Fei;Yin, Fei;Berestetskiy, Alexander

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关键词： destruxin A; interaction; TEME214; SEC23; Bombyx mori

期刊名称：JOURNAL OF FUNGI （ 影响因子：5.816； 五年影响因子：6.499 ）

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年卷期： 2021 年 7 卷 6 期

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收录情况： SCI

摘要： Destruxin A (DA), a mycotoxin isolated from the entomopathogenic fungus Metarhizium anisopliae, has good insecticidal and immune-inhibitory activity, but the action mechanism has not yet been elucidated. In order to identify the DA-binding proteins, we conducted drug affinity responsive target stability (DARTS) experiments, which indicated that the silkworm's (Bombyx mori) transmembrane protein 214 (BmTEME214) and protein transport protein SEC23A isoform X2 (BmSEC23) are the potential DA-binding proteins. The current research was focused on validation of the interaction between DA and these two proteins via bio-layer interferometry (BLI) in vitro, insect two-hybrid (I2H) in Sf9 cells, and RNAi in the insect. The results of the BLI tests showed that DA has strong affinity to bind BmTEME214 and BmSEC23 proteins with a K-D value of 0.286 and 0.291 mu M, respectively. In the I2H experiments, DA inhibited (at 0.02 mu g/mL) and activated (at 0.002-0.0002 mu g/mL) the protein interactions of BmSEC23-BmSEC13, but it only inhibited the BmTMEM214-BmSEC13L interaction. Furthermore, in the RNAi tests, an apparent increase in the silkworm's mortality was recorded in the joint treatment of DA with dsBmSEC23 or dsBmTMEM214 when compared with the single treatment of DA (1.5 mu g/g body), 40 mu g/g body dsBmSEC23, or dsBmTMEM214. This research confirmed that BmSEC23 and BmTMEM214 are the DA-binding proteins and provided new insights to understand the action mechanism of DA.

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