ABSTRACT
Sugars and refined carbohydrates are major components of the modern diet. ATP-citrate lyase (ACLY) is upregulated in adipocytes in response to carbohydrate consumption and generates acetyl-coenzyme A (CoA) for both lipid synthesis and acetylation reactions. Here, we investigate the role of ACLY in the metabolic and transcriptional responses to carbohydrates in adipocytes and unexpectedly uncover a sexually dimorphic function in maintaining systemic metabolic homeostasis. When fed a high-sucrose diet, AclyFAT-/- females exhibit a lipodystrophy-like phenotype, with minimal fat accumulation, insulin resistance, and hepatic lipid accumulation, whereas AclyFAT-/- males have only mild metabolic phenotypes. We find that ACLY is crucial for nutrient-dependent carbohydrate response element-binding protein (ChREBP) activation in adipocytes and plays a key role, particularly in females, in the storage of newly synthesized fatty acids in adipose tissue. The data indicate that adipocyte ACLY is important in females for the systemic handling of dietary carbohydrates and for the preservation of metabolic homeostasis.
Subject(s)
ATP Citrate (pro-S)-Lyase/physiology , Adipocytes/metabolism , Dietary Carbohydrates/administration & dosage , Fatty Acids/metabolism , Homeostasis , Insulin Resistance , Lipogenesis , Acetylation , Adipocytes/cytology , Adult , Animals , Basic Helix-Loop-Helix Leucine Zipper Transcription Factors/metabolism , Female , Humans , Male , Mice , Mice, Knockout , Middle AgedABSTRACT
Cellular metabolism dynamically regulates the epigenome via availability of the metabolite substrates of chromatin-modifying enzymes. The impact of diet on the metabolism-epigenome axis is poorly understood but could alter gene expression and influence metabolic health. ATP citrate-lyase produces acetyl-CoA in the nucleus and cytosol and regulates histone acetylation levels in many cell types. Consumption of a high-fat diet (HFD) results in suppression of ATP citrate-lyase levels in tissues such as adipose and liver, but the impact of diet on acetyl-CoA and histone acetylation in these tissues remains unknown. Here we examined the effects of HFD on levels of acyl-CoAs and histone acetylation in mouse white adipose tissue (WAT), liver, and pancreas. We report that mice consuming a HFD have reduced levels of acetyl-CoA and/or acetyl-CoA:CoA ratio in these tissues. In WAT and the pancreas, HFD also impacted the levels of histone acetylation; in particular, histone H3 lysine 23 acetylation was lower in HFD-fed mice. Genetic deletion of Acly in cultured adipocytes also suppressed acetyl-CoA and histone acetylation levels. In the liver, no significant effects on histone acetylation were observed with a HFD despite lower acetyl-CoA levels. Intriguingly, acetylation of several histone lysines correlated with the acetyl-CoA: (iso)butyryl-CoA ratio in liver. Butyryl-CoA and isobutyryl-CoA interacted with the acetyltransferase P300/CBP-associated factor (PCAF) in liver lysates and inhibited its activity in vitro This study thus provides evidence that diet can impact tissue acyl-CoA and histone acetylation levels and that acetyl-CoA abundance correlates with acetylation of specific histone lysines in WAT but not in the liver.
Subject(s)
Acyl Coenzyme A/metabolism , Adipose Tissue/metabolism , Diet, High-Fat , Histones/metabolism , Liver/metabolism , ATP Citrate (pro-S)-Lyase/genetics , ATP Citrate (pro-S)-Lyase/metabolism , Acetylation , Acyl Coenzyme A/analysis , Animals , Cells, Cultured , Diet, High-Fat/adverse effects , Gene Deletion , Histones/analysis , Male , Mice , Mice, Inbred C57BL , Pancreas/metabolismABSTRACT
Mechanisms of metabolic flexibility enable cells to survive under stressful conditions and can thwart therapeutic responses. Acetyl-coenzyme A (CoA) plays central roles in energy production, lipid metabolism, and epigenomic modifications. Here, we show that, upon genetic deletion of Acly, the gene coding for ATP-citrate lyase (ACLY), cells remain viable and proliferate, although at an impaired rate. In the absence of ACLY, cells upregulate ACSS2 and utilize exogenous acetate to provide acetyl-CoA for de novo lipogenesis (DNL) and histone acetylation. A physiological level of acetate is sufficient for cell viability and abundant acetyl-CoA production, although histone acetylation levels remain low in ACLY-deficient cells unless supplemented with high levels of acetate. ACLY-deficient adipocytes accumulate lipid in vivo, exhibit increased acetyl-CoA and malonyl-CoA production from acetate, and display some differences in fatty acid content and synthesis. Together, these data indicate that engagement of acetate metabolism is a crucial, although partial, mechanism of compensation for ACLY deficiency.
Subject(s)
ATP Citrate (pro-S)-Lyase/metabolism , Acetates/metabolism , Glucose/metabolism , ATP Citrate (pro-S)-Lyase/deficiency , Acetate-CoA Ligase/metabolism , Acetates/pharmacology , Acetyl Coenzyme A/metabolism , Acetylation , Adipocytes/drug effects , Adipocytes/metabolism , Animals , Cell Proliferation/drug effects , Cell Survival/drug effects , Fibroblasts/drug effects , Fibroblasts/metabolism , Gene Deletion , Histones/metabolism , Lipid Metabolism/drug effects , Lipids/biosynthesis , Male , Mice , Up-Regulation/drug effectsABSTRACT
A signaling pathway that rewires metabolism in macrophages to trigger changes in gene expression has been identified.
Subject(s)
Macrophage Activation , Macrophages/metabolism , Gene Expression , Humans , Proto-Oncogene Proteins c-akt/metabolism , Signal TransductionABSTRACT
Salmonellae are pathogenic bacteria that induce immunosuppression by mechanisms that remain largely unknown. Previously, we showed that a putative type II l-asparaginase produced by Salmonella Typhimurium inhibits T cell responses and mediates virulence in a murine model of infection. Here, we report that this putative L-asparaginase exhibits L-asparagine hydrolase activity required for Salmonella Typhimurium to inhibit T cells. We show that L-asparagine is a nutrient important for T cell activation and that L-asparagine deprivation, such as that mediated by the Salmonella Typhimurium L-asparaginase, causes suppression of activation-induced mammalian target of rapamycin signaling, autophagy, Myc expression, and L-lactate secretion. We also show that L-asparagine deprivation mediated by the Salmonella Typhimurium L-asparaginase causes suppression of cellular processes and pathways involved in protein synthesis, metabolism, and immune response. Our results advance knowledge of a mechanism used by Salmonella Typhimurium to inhibit T cell responses and mediate virulence, and provide new insights into the prerequisites of T cell activation. We propose a model in which l-asparagine deprivation inhibits T cell exit from quiescence by causing suppression of activation-induced metabolic reprogramming.
Subject(s)
Asparaginase/physiology , Asparagine/physiology , Bacterial Proteins/physiology , Immune Evasion/physiology , Salmonella typhimurium/enzymology , T-Lymphocyte Subsets/metabolism , Amino Acid Sequence , Animals , Asparaginase/genetics , Asparaginase/pharmacology , Asparagine/deficiency , Asparagine/pharmacology , Autophagy/drug effects , Bacterial Proteins/genetics , Cells, Cultured , Female , Genes, myc , Immune Evasion/genetics , Interleukin-2/biosynthesis , Interleukin-2/genetics , Lactic Acid/metabolism , Lymphocyte Activation , Mice , Mice, Inbred C57BL , Molecular Sequence Data , Proto-Oncogene Proteins c-myc/biosynthesis , Receptors, Antigen, T-Cell, alpha-beta/biosynthesis , Receptors, Antigen, T-Cell, alpha-beta/genetics , Salmonella typhimurium/immunology , Salmonella typhimurium/pathogenicity , T-Lymphocyte Subsets/drug effects , T-Lymphocyte Subsets/immunology , TOR Serine-Threonine Kinases/metabolism , VirulenceABSTRACT
Salmonella enterica serovar Typhimurium avoids clearance by the host immune system by suppressing T cell responses; however, the mechanisms that mediate this immunosuppression remain unknown. We show that S. Typhimurium inhibit T cell responses by producing L-Asparaginase II, which catalyzes the hydrolysis of L-asparagine to aspartic acid and ammonia. L-Asparaginase II is necessary and sufficient to suppress T cell blastogenesis, cytokine production, and proliferation and to downmodulate expression of the T cell receptor. Furthermore, S. Typhimurium-induced inhibition of T cells in vitro is prevented upon addition of L-asparagine. S. Typhimurium lacking the L-Asparaginase II gene (STM3106) are unable to inhibit T cell responses and exhibit attenuated virulence in vivo. L-Asparaginases are used to treat acute lymphoblastic leukemia through mechanisms that likely involve amino acid starvation of leukemic cells, and these findings indicate that pathogens similarly use L-asparagine deprivation to limit T cell responses.