20-Hydroxyecdysone receptor-activated Bombyx mori CCAAT/enhancer-binding protein gamma regulates the expression of BmCBP and subsequent histone H3 lysine 27 acetylation in Bo. mori
20-Hydroxyecdysone receptor-activated Bombyx mori CCAAT/enhancer-binding protein gamma regulates the expression of BmCBP and subsequent histone H3 lysine 27 acetylation in Bo. mori
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20-羟基蜕皮激素受体激活的家蚕 CCAAT/增强子结合蛋白 γ 调节 Bo 中 BmCBP 的表达以及随后的组蛋白 H3 赖氨酸 27 乙酰化。
DOI：
10.1111/imb.12630
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发表时间：
2020-01-06
影响因子：
2.6
通讯作者：
Zheng, S.
中科院分区：
文献类型：
--
作者：
Lyu, H.;Xu, G.;Zheng, S.
环磷酸腺苷（cAMP）反应元件结合蛋白（CREB）-结合蛋白（CBP或CREBBP）在基因转录和动物发育中起着重要的调控作用。然而，CBP被上调以影响昆虫发育的过程是未知的。本研究探讨了20-羟基蜕皮激素（20-hydroxyecdysone，20 E）诱导家蚕CBP（BmCBP）表达的调控机制。在博。mori细胞系DZNU-Bm-12，20 E可增强BmCBP的转录和组蛋白H3 K27的乙酰化。BmCBP RNA干扰（RNAi）导致组蛋白H3 K27乙酰化降低。此外，荧光素酶活性分析显示转录因子Bo. mori CCAAT/增强子结合蛋白γ（BmC/EBPg）激活BmCBP的转录，BmC/EBPg RNAi抑制BmCBP的转录，BmC/EBPg过表达促进BmC/EBPg的转录。电泳迁移率变动分析和染色质免疫沉淀结果表明，BmC/EBPg可以与BmCBP启动子的两个位置的C/EBP顺式调控元件结合。此外，BmC/EBPg的转录增强的20 E受体（BmEcR），结合到BmC/EBPg启动子。BmEcR RNAi显著抑制BmC/EBPg和BmCBP的转录水平。此外，BmEcR-BmC/EBPg途径调节组蛋白H3 K27的乙酰化水平。总之，这些结果表明BmEcR增强BmC/EBPg的表达，BmC/EBPg结合BmCBP启动子，激活BmCBP表达并导致组蛋白H3 K27乙酰化。
Cyclic adenosine monophosphate (cAMP) response element binding protein (CREB)-binding protein (CBP or CREBBP) plays important roles in regulating gene transcription and animal development. However, the process by which CBP is up-regulated to impact insect development is unknown. In this study, the regulatory mechanism of Bombyx mori CBP (BmCBP) expression induced by 20-hydroxyecdysone (20E) was investigated. In the Bo. mori cell line, DZNU-Bm-12, 20E enhanced BmCBP transcription and histone H3K27 acetylation. BmCBP RNA interference (RNAi) resulted in decreased histone H3K27 acetylation. Additionally, the luciferase activity analysis revealed that the transcription factor, Bo. mori CCAAT/enhancer-binding protein gamma (BmC/EBPg), activated BmCBP transcription, which was suppressed by BmC/EBPg RNAi and promoted by BmC/EBPg overexpression. Electrophoretic mobility shift assay and chromatin immunoprecipitation results demonstrated that BmC/EBPg could bind to the C/EBP cis-regulatory elements in two positions of the BmCBP promoter. Moreover, BmC/EBPg transcription was enhanced by the 20E receptor (BmEcR), which bound to the BmC/EBPg promoter. BmEcR RNAi significantly inhibited the transcriptional levels of BmC/EBPg and BmCBP in the presence of 20E. Furthermore, the BmEcR-BmC/EBPg pathway regulated the acetylation levels of histone H3K27. Altogether, these results indicate that BmEcR enhances the expression of BmC/EBPg, which binds to the BmCBP promoter, activates BmCBP expression and leads to histone H3K27 acetylation.