Kruppel homologue 1 interacts directly with Hairy and regulates ecdysis in the brown planthopper
Kruppel homologue 1 interacts directly with Hairy and regulates ecdysis in the brown planthopper
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Krüppel 同源物 1 直接与 Hairy 相互作用并调节褐飞虱的蜕皮
DOI：
10.1111/imb.12635
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发表时间：
2020-01-21
影响因子：
2.6
通讯作者：
Lin, X.
中科院分区：
文献类型：
--
作者：
Mao, Y.;Li, Y.;Lin, X.
幼虫激素（JH）在昆虫的生长发育中起着重要的作用。JH及其受体methoprene- tolerance （Met）通过调控转录因子的表达来控制下游基因的转录。Hairy （Hry）和Kruppel同源物1 （Kr-h1）的表达受JH和JH受体调控。Hry和Kr-h1在介导JH信号传导中都是至关重要的。然而，它们是否在基因水平上相互作用调节变态，以及它们是否在蛋白质水平上物理相互作用仍然未知。我们采用共免疫沉淀、谷胱甘肽s -转移酶下拉和RNA干扰（RNAi）方法研究了两种蛋白Hry和Kr-h1的遗传和生化相互作用。结果表明，褐飞虱Hry与Kr-h1直接相互作用：Hry结合到Kr-h1的n端，其中包含5个锌指结构域。RNAi实验表明，Hry的下调降低了因敲低Kr-h1而导致的电解失败的比例，表明Hry的下调可能通过下调Kr-h1来减轻电解失败。当Kr-h1下调时，Hry的表达显著增加，而当两者均下调时，Hry的表达无显著变化。我们的研究结果表明，Hry蛋白与Kr-h1的结合阻止了n端五个锌指结构域与DNA的结合，从而使Kr-h1的转录激活子或抑制剂功能失活。在未来，Hry可能被用作农药应用的靶标。
Juvenile hormone (JH) plays important roles in the growth and development of insects. JH and its receptor methoprene-tolerant (Met) regulate the expression of transcription factors to control the transcription of downstream genes. The expression of Hairy (Hry) and Kruppel homologue 1 (Kr-h1) is regulated by JH and JH receptors. Hry and Kr-h1 are both crucial in mediating JH signalling. However, whether they interact at the gene level in regulating metamorphosis and whether they interact physically at the protein level remain unknown. We used co-immunoprecipitation, glutathione S-transferase pull-down and RNA interference (RNAi) approaches to study the genetic and biochemical interactions of the two proteins Hry and Kr-h1. The results showed that brown planthopper (Nilaparvata lugens) Hry and Kr-h1 interact directly: Hry binds to the N-terminal of Kr-h1, which includes five zinc-finger domains. The RNAi experiment showed that downregulation of Hry reduced the ratio of ecdysis failure caused by knockdown of Kr-h1, indicating that the downregulation of Hry might mitigate ecdysis failure via the downregulation of Kr-h1. The expression of Hry increased significantly when Kr-h1 was downregulated, whereas it did not change significantly when both were downregulated. Our results suggest that the binding of Hry protein with Kr-h1 prevents the N-terminal five zinc-finger domains from binding with DNA, which in turn inactivates the transcription activator or inhibitor function of Kr-h1. Hry could possibly be used as a target for pesticide applications in the future.