Metabolic and transcriptome responses of RNAi-mediated AMPKα knockdown in Tribolium castaneum

Abstract Background The AMP-activated protein kinase (AMPK) is an intracellular fuel sensor for lipid and glucose metabolism. In addition to the short-term regulation of metabolic enzymes by phosphorylation, AMPK may also exert long-term effects on the transcription of downstream genes through the regulation of transcription factors and coactivators. In this study, RNA interference (RNAi) was conducted to investigate the effects of knockdown of TcAMPKα on lipid and carbohydrate metabolism in the red flour beetle, Tribolium castaneum, and the transcriptome profiles of dsTcAMPKα-injected and dsEGFP-injected beetles under normal conditions were compared by RNA-sequencing.Results RNAi-mediated suppression of TcAMPKα increased whole-body triglyceride (TG) level and the ratio between glucose and trehalose, as was confirmed by in vivo treatment with the AMPK-activating compound, 5-Aminoimidazole-4-carboxamide1-β-D-ribofuranoside (AICAR). A total of 1184 differentially expressed genes (DEGs) were identified between dsTcAMPKα-injected and dsEGFP-injected beetles. These include genes involved in lipid and carbohydrate metabolism as well as insulin/insulin-like growth factor signaling (IIS). Real-time quantitative polymerase chain reaction analysis confirmed the differential expression of selected genes. Interestingly, metabolism-related transcription factors such as sterol regulatory element-binding protein 1 (SREBP1) and carbohydrate response element-binding protein (ChREBP) were also significantly upregulated in dsTcAMPKα-injected beetles. 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.

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Metabolic and transcriptome responses of RNAi-mediated AMPKα knockdown in Tribolium castaneum

10.21203/rs.3.rs-24958/v1 ◽

2020 ◽

Author(s):

Heng Jiang ◽

Nan Zhang ◽

Caihong Ji ◽

Xiangkun Meng ◽

Kun Qian ◽

...

Keyword(s):

Transcription Factors ◽

Carbohydrate Metabolism ◽

Tribolium Castaneum ◽

Binding Protein ◽

Quantitative Polymerase Chain Reaction ◽

Critical Role ◽

Whole Body ◽

Regulation Of Transcription ◽

Valuable Insight ◽

Element Binding Protein

Abstract Background The AMP-activated protein kinase (AMPK) is an intracellular fuel sensor for lipid and glucose metabolism. In addition to the short-term regulation of metabolic enzymes by phosphorylation, AMPK may also exert long-term effects on the transcription of downstream genes through the regulation of transcription factors and coactivators. In this study, RNA interference (RNAi) was conducted to investigate the effects of knockdown of TcAMPKα on lipid and carbohydrate metabolism in the red flour beetle, Tribolium castaneum, and the transcriptome profiles of dsTcAMPKα-injected and dsEGFP-injected beetles under normal conditions were comparatively analyzed by RNA-sequencing. Results RNAi-mediated suppression of TcAMPKα increased whole-body triglyceride (TG) level and the ratio between glucose and trehalose, as was confirmed by in vivo treatment with the AMPK-activating compound, 5-Aminoimidazole-4-carboxamide1-β-D-ribofuranoside (AICAR). A total of 1184 differentially expressed genes (DEGs) were identified between dsTcAMPKα-injected and dsEGFP-injected beetles. These include genes involved in lipid and carbohydrate metabolism as well as insulin/insulin-like growth factor signaling (IIS). Real-time quantitative polymerase chain reaction analysis confirmed the differential expression of selected genes. Interestingly, metabolism-related transcription factors such as sterol regulatory element-binding protein 1 (SREBP1) and carbohydrate response element-binding protein (ChREBP) were also significantly upregulated in dsTcAMPKα-injected beetles. 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.

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Metabolic and transcriptome responses of RNAi-mediated AMPKα knockdown in Tribolium castaneum

BMC Genomics ◽

10.1186/s12864-020-07070-3 ◽

2020 ◽

Vol 21 (1) ◽

Author(s):

Heng Jiang ◽

Nan Zhang ◽

Caihong Ji ◽

Xiangkun Meng ◽

Kun Qian ◽

...

Keyword(s):

Transcription Factors ◽

Carbohydrate Metabolism ◽

Tribolium Castaneum ◽

Binding Protein ◽

Quantitative Polymerase Chain Reaction ◽

Critical Role ◽

Whole Body ◽

Regulation Of Transcription ◽

Valuable Insight ◽

Element Binding Protein

Abstract Background The AMP-activated protein kinase (AMPK) is an intracellular fuel sensor for lipid and glucose metabolism. In addition to the short-term regulation of metabolic enzymes by phosphorylation, AMPK may also exert long-term effects on the transcription of downstream genes through the regulation of transcription factors and coactivators. In this study, RNA interference (RNAi) was conducted to investigate the effects of knockdown of TcAMPKα on lipid and carbohydrate metabolism in the red flour beetle, Tribolium castaneum, and the transcriptome profiles of dsTcAMPKα-injected and dsEGFP-injected beetles under normal conditions were compared by RNA-sequencing. Results RNAi-mediated suppression of TcAMPKα increased whole-body triglyceride (TG) level and the ratio between glucose and trehalose, as was confirmed by in vivo treatment with the AMPK-activating compound, 5-Aminoimidazole-4-carboxamide1-β-D-ribofuranoside (AICAR). A total of 1184 differentially expressed genes (DEGs) were identified between dsTcAMPKα-injected and dsEGFP-injected beetles. These include genes involved in lipid and carbohydrate metabolism as well as insulin/insulin-like growth factor signaling (IIS). Real-time quantitative polymerase chain reaction analysis confirmed the differential expression of selected genes. Interestingly, metabolism-related transcription factors such as sterol regulatory element-binding protein 1 (SREBP1) and carbohydrate response element-binding protein (ChREBP) were also significantly upregulated in dsTcAMPKα-injected beetles. 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.

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Metabolic and transcriptome responses of RNAi-mediated AMPKα knockdown in Tribolium castaneum

10.21203/rs.3.rs-24958/v2 ◽

2020 ◽

Author(s):

Heng Jiang ◽

Nan Zhang ◽

Caihong Ji ◽

Xiangkun Meng ◽

Kun Qian ◽

...

Keyword(s):

Transcription Factors ◽

Carbohydrate Metabolism ◽

Tribolium Castaneum ◽

Binding Protein ◽

Quantitative Polymerase Chain Reaction ◽

Critical Role ◽

Whole Body ◽

Regulation Of Transcription ◽

Valuable Insight ◽

Element Binding Protein

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New perspectives in the regulation of hepatic glycolytic and lipogenic genes by insulin and glucose: a role for the transcription factor sterol regulatory element binding protein-1c

Biochemical Journal ◽

10.1042/bj20020430 ◽

2002 ◽

Vol 366 (2) ◽

pp. 377-391 ◽

Cited By ~ 323

Author(s):

Fabienne FOUFELLE ◽

Pascal FERRÉ

Keyword(s):

Gene Expression ◽

Transcription Factors ◽

Binding Protein ◽

Regulatory Element ◽

Fine Tuning ◽

Whole Body ◽

Transcriptional Level ◽

Hepatic Glucose ◽

Element Binding Protein ◽

Sterol Regulatory Element

The regulation of hepatic glucose metabolism has a key role in whole-body energy metabolism, since the liver is able to store (glycogen synthesis, lipogenesis) and to produce (glycogenolysis, gluconeogenesis) glucose. These pathways are regulated at several levels, including a transcriptional level, since many of the metabolism-related genes are expressed according to the quantity and quality of nutrients. Recent advances have been made in the understanding of the regulation of hepatic glycolytic, lipogenic and gluconeogenic gene expression by pancreatic hormones, insulin and glucagon and glucose. Here we review the role of the transcription factors forkhead and sterol regulatory element binding protein-1c in the inductive and repressive effects of insulin on hepatic gene expression, and the pathway that leads from glucose to gene regulation with the recently discovered carbohydrate response element binding protein. We discuss how these transcription factors are integrated in a regulatory network that allows a fine tuning of hepatic glucose storage or production, and their potential importance in metabolic diseases.

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Study of expression analysis of SIRT4 and the coordinate regulation of bovine adipocyte differentiation by SIRT4 and its transcription factors

Bioscience Reports ◽

10.1042/bsr20181705 ◽

2018 ◽

Vol 38 (6) ◽

Cited By ~ 6

Author(s):

Jieyun Hong ◽

Shijun Li ◽

Xiaoyu Wang ◽

Chugang Mei ◽

Linsen Zan

Keyword(s):

Transcriptional Regulation ◽

Transcription Factors ◽

Adipocyte Differentiation ◽

Binding Protein ◽

Core Promoter ◽

Critical Role ◽

Luciferase Reporter ◽

Marker Genes ◽

Gene Assay ◽

Bovine Adipocytes

Sirtuins, NAD+-dependent deacylases and ADP-ribosyltransferases, are critical regulators of metabolism involved in many biological processes, and are involved in mediating adaptive responses to the cellular environment. SIRT4 is a mitochondrial sirtuin and has been shown to play a critical role in maintaining insulin secretion and glucose homeostasis. As a regulator of lipid homeostasis, SIRT4 can repress fatty acid oxidation and promote lipid anabolism in nutrient-replete conditions. Using real-time quantitative PCR (qPCR) to explore the molecular mechanisms of transcriptional regulation of bovine SIRT4 during adipocyte differentiation, we found that bovine SIRT4 is expressed at high levels in bovine subcutaneous adipose tissue. SIRT4 knockdown led to decreased expression of adipogenic differentiation marker genes during adipocyte differentiation. The core promoter of bovine SIRT4 was identified in the −402/−60 bp region of the cloned 2-kb fragment containing the 5′-regulatory region. Binding sites were identified in this region for E2F transcription factor-1 (E2F1), CCAAT/enhancer-binding protein β (CEBPβ), homeobox A5 (HOXA5), interferon regulatory factor 4 (IRF4), paired box 4 (PAX4), and cAMP responsive element-binding protein 1 (CREB1) by using Electrophoretic mobility shift assay (EMSA) and luciferase reporter gene assay. We also found that E2F1, CEBPβ, and HOXA5 transcriptionally activate SIRT4 expression, whereas, IRF4, PAX4, and CREB1 transcriptionally repress SIRT4 expression. We further verified that SIRT4 knockdown could affect the ability of these transcription factors (TFs) to regulate the differentiation of bovine adipocytes. In conclusion, our results shed light on the mechanisms underlying the transcriptional regulation of SIRT4 expression in bovine adipocytes.

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RNA-Binding Protein MSI2 Binds to miR-301a-3p and Facilitates Its Distribution in Mitochondria of Endothelial Cells

Frontiers in Molecular Biosciences ◽

10.3389/fmolb.2020.609828 ◽

2021 ◽

Vol 7 ◽

Author(s):

Qian Qian Guo ◽

Jing Gao ◽

Xiao Wei Wang ◽

Xian Lun Yin ◽

Shu Cui Zhang ◽

...

Keyword(s):

Endothelial Cells ◽

Rna Binding ◽

Binding Protein ◽

Umbilical Vein ◽

Quantitative Polymerase Chain Reaction ◽

Rna Binding Protein ◽

Small Rna Sequencing ◽

Valuable Insight ◽

Label Free ◽

Mitochondrial Distribution

Numerous miRNAs have been detected in mitochondria, which play important roles in many physiological and pathophysiological processes. However, the dynamic changes of miRNA distribution in mitochondria and their mechanisms in reactive oxygen species (ROS)-induced endothelial injury remain unclear. Therefore, miRNA levels in whole cells and mitochondria of H2O2-treated endothelial cells were analyzed by small RNA sequencing in the present study. The results showed that H2O2 significantly reduced the relative mitochondrial distribution of dozens of miRNAs in human umbilical vein endothelial cells (HUVECs). Among the high-abundance miRNAs, miR-301a-3p has the most significant changes in the redistribution between cytosol and mitochondria confirmed by absolute quantitative polymerase chain reaction (qPCR). To unravel the mechanism of miR-301a-3p distribution in mitochondria, RNA pull-down followed by label-free quantitative proteomic analysis was performed, and RNA-binding protein Musashi RNA binding protein 2 (MSI2) was found to specifically bind to miR-301a-3p. Western blotting and immunofluorescence colocalization assay showed that MSI2 was located in mitochondria of various cell types. H2O2 significantly downregulated MSI2 expression in whole endothelial cells, promoted the distribution of MSI2 in cytosol and decreased its distribution in the mitochondria. Moreover, overexpression of MSI2 increased the mitochondrial distribution of miR-301a-3p, whereas inhibition of MSI2 decreased its distribution in mitochondria. Thus, MSI2 might be responsible for the distribution of miR-301a-3p between cytosol and mitochondria in endothelial cells. Our findings revealed for the first time that MSI2 was involved in the regulation of miRNA distribution in mitochondria and provided valuable insight into the mechanism of mitochondrial distribution of miRNAs.

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Modification of DNA topoisomerase II activity via direct interactions with the cyclic adenosine-3',5'-monophosphate response element-binding protein and related transcription factors.

Molecular Endocrinology ◽

10.1210/mend.7.3.8387155 ◽

1993 ◽

Vol 7 (3) ◽

pp. 305-318 ◽

Cited By ~ 1

Author(s):

D J Kroll ◽

D M Sullivan ◽

A Gutierrez-Hartmann ◽

J P Hoeffler

Keyword(s):

Transcription Factors ◽

Topoisomerase Ii ◽

Binding Protein ◽

Dna Topoisomerase Ii ◽

Dna Topoisomerase ◽

Response Element ◽

Element Binding Protein

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A New Role for Sterol Regulatory Element Binding Protein 1 Transcription Factors in the Regulation of Muscle Mass and Muscle Cell Differentiation

Molecular and Cellular Biology ◽

10.1128/mcb.00690-09 ◽

2009 ◽

Vol 30 (5) ◽

pp. 1182-1198 ◽

Cited By ~ 54

Author(s):

Virginie Lecomte ◽

Emmanuelle Meugnier ◽

Vanessa Euthine ◽

Christine Durand ◽

Damien Freyssenet ◽

...

Keyword(s):

Transcription Factors ◽

Muscle Mass ◽

Target Genes ◽

Binding Protein ◽

Regulatory Element ◽

Element Binding Protein ◽

Sterol Regulatory Element ◽

Myogenic Program

ABSTRACT The role of the transcription factors sterol regulatory element binding protein 1a (SREBP-1a) and SREBP-1c in the regulation of cholesterol and fatty acid metabolism has been well studied; however, little is known about their specific function in muscle. In the present study, analysis of recent microarray data from muscle cells overexpressing SREBP1 suggested that they may play a role in the regulation of myogenesis. We then demonstrated that SREBP-1a and -1c inhibit myoblast-to-myotube differentiation and also induce in vivo and in vitro muscle atrophy. Furthermore, we have identified the transcriptional repressors BHLHB2 and BHLHB3 as mediators of these effects of SREBP-1a and -1c in muscle. Both repressors are SREBP-1 target genes, and they affect the expression of numerous genes involved in the myogenic program. Our findings identify a new role for SREBP-1 transcription factors in muscle, thus linking the control of muscle mass to metabolic pathways.

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Nuclear Factor of Activated T Cells (NFAT)-dependent Transactivation Regulated by the Coactivators p300/CREB-binding Protein (CBP)

Journal of Experimental Medicine ◽

10.1084/jem.187.12.2031 ◽

1998 ◽

Vol 187 (12) ◽

pp. 2031-2036 ◽

Cited By ~ 134

Author(s):

Carmen García-Rodríguez ◽

Anjana Rao

Keyword(s):

T Cells ◽

Binding Protein ◽

Critical Role ◽

Regulation Of Transcription ◽

Dependent Manner ◽

Nuclear Factor ◽

Transactivation Domain ◽

Creb Binding Protein ◽

Activated T Cells ◽

Viral Oncoprotein

p300 and cAMP response element–binding protein (CREB)–binding protein (CBP) are members of a family of coactivators involved in the regulation of transcription and chromatin. We show that transcription factors of the nuclear factor of activated T cells (NFAT) family bind p300/CBP and recruit histone acetyltransferase activity from T cell nuclear extracts. The NH2-terminal transactivation domain of NFAT1 and the phospho-CREB- and E1A-binding sites of p300/CBP are involved in the interaction. The viral oncoprotein E1A inhibits NFAT-dependent transactivation in a p300-dependent manner. Recruitment of the coactivators p300/CBP by the transactivation domains of NFAT proteins is likely to play a critical role in NFAT-dependent gene expression during the immune response.

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