The Direct Interaction between E93 and Kr-h1 Mediated Their Antagonistic Effect on Ovary Development of the Brown Planthopper
The Direct Interaction between E93 and Kr-h1 Mediated Their Antagonistic Effect on Ovary Development of the Brown Planthopper
复制标题
E93与Kr-h1直接相互作用介导对褐飞虱卵巢发育的拮抗作用
DOI：
10.3390/ijms20102431
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发表时间：
2019-05-02
影响因子：
5.6
通讯作者：
Lin, Xinda
中科院分区：
文献类型：
--
作者：
Mao, Yiwen;Li, Yan;Lin, Xinda
保幼激素(JH)信号通路和蜕皮激素信号通路是协调昆虫变态的重要内分泌信号通路。变态过程，即从未成熟形态到成体形态的变化，是由JH和下游转录因子的急剧减少所指挥的。Krüppel同系物1(Kr-H1)是JH信号通路的下游转录因子，具有抗变性作用，抑制e93的表达。然而，Kr-H1和e93之间的生化相互作用以及这种相互作用如何调节卵巢发育--内分泌调节的灵敏读数--仍然未知。在褐飞虱中，我们发现Kr-H1基因的下调部分恢复了e93基因敲除对变态的恶化作用。与单独敲除E93相比，双重敲除E93和Kr-H1基因可增加卵巢发育和产卵量，表明Kr-H1基因敲除部分恢复了e93敲除对卵巢发育的恶化作用。综上所述，我们的结果表明，E93和Kr-H1在调节变态和卵巢发育方面具有拮抗作用。我们测试了这两种蛋白质之间的生化相互作用，发现这些分子直接相互作用。KR-H1 V和E93 II发生了强烈而特异的相互作用，表明潜在的相互作用结构域可能位于这两个区域。我们推测，PinSqueak家族的核受体相互作用基序(NR-box)和螺旋-转弯-螺旋DNA结合基序(RHF1)是负责E93和Kr-H1之间蛋白质-蛋白质相互作用的候选结构域。此外，HA标记的e93和标志标记的Kr-H1共定位于细胞核内，当Kr-H1表达下调时，e93的表达增加，支持这两种蛋白可能存在拮抗作用。JH和蜕皮激素信号对控制卵巢发育和害虫种群是至关重要的。我们的结果对于理解e93和相关蛋白质之间的相互作用很重要，这使得识别潜在的靶标和开发新的杀虫剂用于害虫管理成为可能。
The juvenile hormone (JH) signalling and ecdysone signalling pathways are crucial endocrine signalling pathways that orchestrate the metamorphosis of insects. The metamorphic process, the morphological change from the immature to adult forms, is orchestrated by the dramatic reduction of JH and downstream transcription factors. The Krüppel-homologue 1 (Kr-h1), a downstream transcription factor of the JH signalling pathway, represses E93 expression with an anti-metamorphic effect. However, the biochemical interaction between Kr-h1 and E93 and how the interaction regulates ovary development, a sensitive readout for endocrine regulation, remain unknown. In brown planthopper, Nilaparvata lugens, we found that the downregulation of Kr-h1 partially recovered the deteriorating effect of E93 knock-down on metamorphosis. Dual knock down of E93 and Kr-h1 increased ovary development and the number of eggs laid when compared to the effects of the knock down of E93 alone, indicating that the knock down of Kr-h1 partially recovered the deteriorating effect of the E93 knock-down on ovary development. In summary, our results indicated that E93 and Kr-h1 have antagonistic effects on regulating metamorphosis and ovary development. We tested the biochemical interaction between these two proteins and found that these molecules interact directly. Kr-h1 V and E93 II undergo strong and specific interactions, indicating that the potential interacting domain may be located in these two regions. We inferred that the nuclear receptor interaction motif (NR-box) and helix-turn-helix DNA binding motifs of the pipsqueak family (RHF1) are candidate domains responsible for the protein–protein interaction between E93 and Kr-h1. Moreover, the HA-tagged E93 and FLAG-tagged Kr-h1 were co-localized in the nucleus, and the expression of E93 was increased when Kr-h1 was downregulated, supporting that these two proteins may interact antagonistically. JH and ecdysone signalling are critical for the control of ovary development and pest populations. Our result is important for understanding the interactions between E93 and related proteins, which makes it possible to identify potential targets and develop new pesticides for pest management.