- Background: The AMP-activated protein kinase (AMPK) is an intracellular fuel sensor for lipid and glucose metabolism. - In this study, RNA interference (RNAi) was conducted to investigate the effects of knockdown of TcAMPK α on lipid and carbohydrate metabolism in the red flour beetle, Tribolium castaneum, and the transcriptome profiles of dsTcAMPK α -injected and dsEGFP-injected beetles under normal conditions were compared by RNA- sequencing.. - Conclusions: AMPK plays a critical role in the regulation of beetle metabolism. - The findings of DEGs involved in lipid and carbohydrate metabolism provide valuable insight into the role of AMPK signaling in the transcriptional regulation of insect metabolism.. - The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. - If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. - AMP-activated protein kin- ase (AMPK) is a cellular energy sensor conserved across all eukaryotic species [13]. - As a serine/threonine protein kinase complex, AMPK consists of a catalytic subunit α and two regulatory subunits, β and γ, and is activated in response to energy stress by sensing increases in ADP/. - While nucleotide- dependent phosphorylation of Thr172 in the α subunit by liver kinase B1 (LKB1) is the principal event required for full activation of AMPK in mammalian cells [19, 70], several studies have revealed the nucleotide-independent regulation of AMPK via the phosphorylation of Thr172 by calcium/calmodulin-dependent kinase kinase 2 in mammals (CAMKK . - Although the role of AMPK in the regulation of cell metabolism is well studied in mammals, related research is still limited in insects. - Notably, AMPK can regulate energy balance via modulation of transcriptional expres- sion of metabolic enzymes in the long term, however, its downstream transcriptional pathways remains largely elusive [7]. - Recently, we reported the transcriptional and post-translational activation of TcAMPKα by oxidative, heat and cold stresses in the red flour beetle, Tribolium castaneum [25]. - 8.61% (ANOVA, df 1, 4 , F = 7.917, P value when compared with the dsEGFP group mmol/mgprot) (Fig. - (ANOVA, df 1, 4 , F = 8.196, P value was ob- served in the beetles injected with dsTcAMPKα μmol/g) when compared to the control beetles μmol/g) (Fig. - 3.01% (ANOVA, df 1, 4 , F = 9.357, P value = 0.0377) than that in dsEGFP group mg/g) (Fig. - (ANOVA, df 1, 4 , F = 10.770, P value and 41.89. - 2.27% (ANOVA, df 1, 4 , F = 93.320, P value respectively, compared with injection buffer (IB). - 4.02% (ANOVA, df 1, 4 , F = 8.910, P value = 0.0405) in beetles treated with AICAR. - cas- taneum showed that 83.50 and 77.15% reads of the dsEGFP and dsTcAMPKα groups were aligned on aver- age, respectively (Table 2).. - Similarly, among the 42 DEGs in the post- translational modification class, 35 DEGs were down- regulated, including 3 heat shock proteins (Hsps), whereas the phosphatidylinositol 4,5-bisphosphate 3- kinase (PIK3) and InR2 involved in the signal transduc- tion mechanisms were upregulated.. - Besides the analysis of the entire gene set, we specifically checked for up- or downregulation of lipid and carbohy- drate metabolism genes in our set of DEGs. - ANOVA, df 1, 4 , F = 5.152, P value . - TC015337 and TC015340) (Fold change: 2.03 and 3.22, ANOVA, df 1, 4 , F = 9.182 and 157.306, P value = 0.03878 and 0.00023) involved in fatty acid biosynthetic pathways, and the transcription factor ChREBP (Accession no.. - TC011996, TC034013 and TC010784) (Fold change and 2.46. - ANOVA, df 1, 4 , F and 34.155, P value and . - To confirm the reliability of the DEG Table 1 Summary of the transcriptome sequencing data from. - TC011522, TC015400 and TC000238) (Fold change and 1.50;. - ANOVA, df 1, 4 , F and 2.781, P value and 0.17070), one ACC (Accession no. - TC015612) (Fold change: 1.77. - ANOVA, df 1, 4 , F. - 3 Correlation and box plot of the gene expression levels (FPKM) for all of the samples. - A: Pearson correlations of gene expression levels of the six samples. - B: Box plots of gene expression levels of the six samples.. - X-axis: log 2 -fold change (treatment/control). - TC004512) (Fold change: 1.59. - ANOVA, df 1, 4 , F = 8.092, P value four trehalase (TRE) genes responsible for trehalose hydrolysis (Accession no. - TC006698, LOC659620, TC004791 and TC006697) (Fold change and 1.86. - ANOVA, df 1, 4 , F and 9.491, P value and 0.03690), two genes involved in gluconeogenesis including pyruvate carboxylase (PC) (Accession no. - TC032730) (Fold change: 1.73. - ANOVA, df 1, 4 , F = 19.919, P value and phosphoenolpyr- uvate carboxykinase (PEPCK) (Accession no. - TC009072) (Fold change: 1.23. - ANOVA, df 1, 4 , F = 1.491, P value were significantly upregulated (Table 4). - TC007163) (Fold change: 1.93;. - ANOVA, df 1, 4 , F = 9.490, P value the master regulator of lipid homeostasis, and SREBP cleavage- activating protein (SCAP) (Accession no. - TC013456) (Fold change: 1.75. - These results were further confirmed by in vivo AICAR treatment, which resulted in the decreased TG and glucose levels and increased trehalose content. - On the other hand, while activation of AMPK triggered a. - 5 KOG function classifications of the differentially expressed unigenes. - The X-axis represents names of 25 groups, and the Y-axis corresponds to the number of unigenes in the group. - These results indicate the complexity of the role of AMPK in the regulation of metabolic processes.. - We also observed the upregula- tion of several carbohydrate metabolism-related genes, such as TRE, and two key enzymes in gluco- neogenesis (de novo synthesis of glucose), PC and PEPCK [16, 26], in the dsTcAMPKα-injected insects.. - SNF1A/AMP-activated protein kinase;. - Fatty acid synthetase. - FAS fatty acid synthase . - IIS pathway was involved in the regulation of glucose and lipid metabolism (Saltie and Kahn 2001) [50]. - In this study, we observed the up- regulation of IIS-related DEGs like IRS1, InR2 and PI3K in dsTcAMPKα-injected beetles, which might result in the upregulation of SREBP1 and ChREBP. - This study confirmed that AMPK has an important role in the regulation of beetle metabolism. - 0.05 and fold change |log 2 (treatment/control. - AMPK: AMP-activated protein kinase. - FAS: fatty acid synthetase. - The funding bodies played no role in the design of the study and collection, analysis, and interpretation of data and in writing the manuscript.. - Chitin synthases are required for survival, fecundity and egg hatch in the red flour beetle, Tribolium castaneum. - AMP-activated protein kinase and its downstream transcriptional pathways. - A common bicyclic protein kinase cascade inactivates the regulatory enzymes of fatty acid and cholesterol biosynthesis. - The regulation of AMP-activated protein kinase by H 2 O 2 . - Phosphorylation of the insulin receptor by AMP-activated protein kinase (AMPK) promotes ligand-independent activation of the insulin signalling pathway in rodent muscle. - Status of malathion resistance in five genera of beetles infesting farm-stored corn, wheat, and oats in the United States. - Regulation of phosphoenolpyruvate carboxykinase (GTP) gene expression. - AMP-activated/SNF1 protein kinases: conserved guardians of cellular energy. - Roles of the Snf1/Rkin1/AMP-activated protein kinase family in the response to environmental and nutritional stress. - Complexes between the LKB1 tumor suppressor, STRAD α / β and MO25 α / β are upstream kinases in the AMP-activated protein kinase cascade. - Calmodulin-dependent protein kinase kinase- β is an alternative upstream kinase for AMP-activated protein kinase. - The Ca 2+ /calmodulin-dependent protein kinase kinases are AMP-activated protein kinase kinases. - 5 ′ -AMP-activated protein kinase phosphorylates IRS-1 on Ser-789 in mouse C2C12 myotubes in response to 5-aminoimidazole-4-carboxamide riboside. - Transcriptional and post-translational activation of AMPK α by oxidative, heat, and cold stresses in the red flour beetle, Tribolium castaneum. - Mechanism for fatty acid “ sparing ” effect on glucose-induced transcription regulation of carbohydrate-responsive element-binding protein by AMP-activated protein kinase. - dephosphorylation of the carbohydrate response element binding protein.. - Differential regulation of insulin receptor substrates-1 and-2 (IRS-1 and IRS-2) and phosphatidylinositol 3- kinase isoforms in liver and muscle of the obese diabetic (Ob/Ob) mouse. - Functions of duplicated genes encoding CCAP receptors in the red flour beetle, Tribolium castaneum. - The stimulation of glycolysis by hypoxia in activated monocytes is mediated by AMP-activated protein kinase and inducible 6-phosphofructo-2-kinase. - Phosphorylation and activation of heart PFK-2 by AMPK has a role in the stimulation of glycolysis during ischaemia. - Histochemical and ultrastructural studies of the mosquito Aedes aegypti fat body: effects of aging and diet type. - AMP-activated kinase reciprocally regulates triacylglycerol synthesis and fatty acid oxidation in. - Targeting the AMP-activated protein kinase for the treatment of type 2 diabetes. - The transcriptional response of the yeast Na + -ATPase ENA1 gene to alkaline stress involves three main signaling pathways. - AMP-activated protein kinase and metabolic regulation in cold-hardy insects. - Insulin signalling and the regulation of glucose and lipid metabolism. - AMP-activated protein kinase (AMPK) control of fatty acid and glucose metabolism in the ischemic heart. - IGF-1 induces SREBP-1 expression and lipogenesis in SEB-1 sebocytes via activation of the phosphoinositide 3-kinase/Akt pathway. - Complementary roles of IRS-1 and IRS-2 in the hepatic regulation of metabolism. - AMP-activated protein kinase in metabolic control and insulin signaling. - Glucose repression/derepression in budding yeast: SNF1 protein kinase is activated by phosphorylation under derepressing conditions, and this correlates with a high AMP: ATP ratio. - Inactivation of acetyl-CoA carboxylase and activation of AMP-activated protein kinase in muscle during exercise. - Ca 2+ /calmodulin-dependent protein kinase kinase- β acts upstream of AMP-activated protein kinase in mammalian cells. - LKB1 is the upstream kinase in the AMP-activated protein kinase cascade. - Juvenile hormone and insulin regulate trehalose homeostasis in the red flour beetle, Tribolium castaneum. - Adiponectin stimulates glucose utilization and fatty- acid oxidation by activating AMP-activated protein kinase. - Hypothalamic AMP-activated protein kinase regulates glucose production. - AMP-activated protein kinase is required for the lipid-lowering effect of metformin in insulin-resistant human HepG2 cells. - Role of AMP-activated protein kinase in mechanism of metformin action
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