title: The steroid hormone 20-hydroxyecdysone via nongenomic pathway activates Ca2+/calmodulin-dependent protein kinase II to regulate gene expression.
abstract: The steroid hormone 20-hydroxyecdysone (20E) triggers calcium signaling pathway to regulate 20E response gene expression, but the mechanism underlying this process remains unclear. We propose that the 20E-induced phosphorylation of Ca(2+)/calmodulin-dependent protein kinase II (CaMKII) serves an important function in 20E response gene transcription in the lepidopteran insect Helicoverpa armigera. CaMKII showed increased expression and phosphorylation during metamorphosis. 20E elevated CaMKII phosphorylation. However, the G protein-coupled receptor (GPCR) and ryanodine receptor inhibitor suramin, the phospholipase C inhibitor U73122, and the inositol 1,4,5-triphosphate receptor inhibitor xestospongin C suppressed 20E-induced CaMKII phosphorylation. Two ecdysone-responsible GPCRs and Gαq protein were involved in 20E-induced CaMKII phosphorylation by RNA interference analysis. 20E regulated CaMKII threonine phosphorylation at amino acid 290, thereby inducing CaMKII nuclear translocation. CaMKII knockdown by dsCaMKII injection into the larvae prevented the occurrence of larval-pupal transition and suppressed 20E response gene expression. CaMKII phosphorylation and nuclear translocation maintained USP1 lysine acetylation at amino acid 303 by inducing histone deacetylase 3 phosphorylation and nuclear export. The lysine acetylation of USP1 was necessary for the interaction of USP1 with EcRB1 and their binding to the ecdysone response element. Results suggest that 20E (via GPCR activation and calcium signaling) activates CaMKII phosphorylation and nuclear translocation, which regulate USP1 lysine acetylation to form an EcRB1-USP1 complex for 20E response gene transcription.
authors: From the Shandong Provincial Key Laboratory of Animal Cells and Developmental Biology, School of Life Sciences, Shandong University, Jinan 250100, China.
authors: Yu-Pu
authors: YP
authors: Jing
authors: From the Shandong Provincial Key Laboratory of Animal Cells and Developmental Biology, School of Life Sciences, Shandong University, Jinan 250100, China.
authors: Wen
authors: W
authors: Liu
authors: From the Shandong Provincial Key Laboratory of Animal Cells and Developmental Biology, School of Life Sciences, Shandong University, Jinan 250100, China.
authors: Jin-Xing
authors: JX
authors: Wang
authors: From the Shandong Provincial Key Laboratory of Animal Cells and Developmental Biology, School of Life Sciences, Shandong University, Jinan 250100, China xfzhao@sdu.edu.cn.
authors: Xiao-Fan
authors: XF
authors: Zhao
mesh_terms: D000107:Acetylation; D021581:Active Transport, Cell Nucleus; D000595:Amino Acid Sequence; D000818:Animals; D054732:Calcium-Calmodulin-Dependent Protein Kinase Type 2 / Q000378:metabolism*; D002460:Cell Line; D002467:Cell Nucleus / Q000201:enzymology; D004441:Ecdysterone / Q000502:physiology*; D004789:Enzyme Activation; D044127:Epigenesis, Genetic; D018507:Gene Expression Regulation, Developmental; D006655:Histone Deacetylases / Q000378:metabolism; D019476:Insect Proteins / Q000378:metabolism*; D008675:Metamorphosis, Biological; D008969:Molecular Sequence Data; D009036:Moths / Q000201:enzymology / Q000235:genetics* / Q000254:growth & development; D010766:Phosphorylation; D011485:Protein Binding; D011499:Protein Processing, Post-Translational; D020218:Response Elements; D064570:Ubiquitin-Specific Proteases / Q000378:metabolism; D000099186:Histone Deacetylase 3
chemical_list: D054732:Calcium-Calmodulin-Dependent Protein Kinase Type 2; D004441:Ecdysterone; D006655:Histone Deacetylases; D019476:Insect Proteins; D064570:Ubiquitin-Specific Proteases; D000099186:Histone Deacetylase 3
keywords: 20-Hydroxyecdysone; Ca2+/Calmodulin-dependent Protein Kinase II (CaMKII); Histone Deacetylase 3 (HDAC3); Phosphorylation; Steroid Hormone; Transcription Regulation; USP1 Lysine Acetylation