Saturated Very Long Chain Fatty Acids Are Required for the Production of Infectious Human Cytomegalovirus Progeny

Koyuncu, Emre; Purdy, John; Rabinowitz, Joshua D.; Shenk, Thomas

doi:10.1371/journal.ppat.1003333

Cited by 94 publications

(134 citation statements)

References 60 publications

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“…Infectious supernatant was applied to uninfected fibroblasts, and 24-36 h later, cells were fixed with 4% (wt/vol) paraformaldehyde and immunostained for IE1 (19,66). For assaying cell-associated viral spread, infected ARPE-19 cells were directly fixed with 4% (wt/vol) paraformaldehyde at 8.5 dpi and immunostained for IE1.…”

Section: Methodsmentioning

confidence: 99%

“…Cortisol, the glucocorticoid receptor ligand, enhanced virus production in human embryonic kidney cells (16). Most recently, a PPAR response element was described within the MIEP, and a PPARγ antagonist reduced viral protein expression, lipid droplet formation, and virus production (17).To replicate efficiently, HCMV induces numerous metabolic and lipidomic changes (18,19), and the intimate connection between metabolism and nuclear receptors (20, 21) offers a compelling reason to examine the role of these transcription factors during infection more closely. In addition, nuclear receptors are also known to modulate inflammatory signaling (22) and cell cycle progression (23), pathways that are important for successful infection.…”

mentioning

confidence: 99%

“…To replicate efficiently, HCMV induces numerous metabolic and lipidomic changes (18,19), and the intimate connection between metabolism and nuclear receptors (20, 21) offers a compelling reason to examine the role of these transcription factors during infection more closely. In addition, nuclear receptors are also known to modulate inflammatory signaling (22) and cell cycle progression (23), pathways that are important for successful infection.…”

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confidence: 99%

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Estrogen-related receptor α is required for efficient human cytomegalovirus replication

Hwang¹,

Purdy

Wu³

et al. 2014

Proc. Natl. Acad. Sci. U.S.A.

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An shRNA-mediated screen of the 48 human nuclear receptor genes identified multiple candidates likely to influence the production of human cytomegalovirus in cultured human fibroblasts, including the estrogen-related receptor α (ERRα), an orphan nuclear receptor. The 50-kDa receptor and a 76-kDa variant were induced posttranscriptionally following infection. Genetic and pharmacological suppression of the receptor reduced viral RNA, protein, and DNA accumulation, as well as the yield of infectious progeny. In addition, RNAs encoding multiple metabolic enzymes, including enzymes sponsoring glycolysis (enolase 1, triosephosphate isomerase 1, and hexokinase 2), were reduced when the function of ERRα was inhibited in infected cells. Consistent with the effect on RNAs, a substantial number of metabolites, which are normally induced by infection, were either not increased or were increased to a reduced extent in the absence of normal ERRα activity. We conclude that ERRα is needed for the efficient production of cytomegalovirus progeny, and we propose that the nuclear receptor contributes importantly to the induction of a metabolic environment that supports optimal cytomegalovirus replication.herpesvirus | metabolism | nuclear receptor | mass spectrometry H uman cytomegalovirus (HCMV) belongs to the β-herpesvirus subfamily, and although most healthy individuals remain asymptomatic subsequent to infection, the pathogen is a major contributor to birth defects and to life-threatening disease in immunocompromised patients (1-3). As with all viruses, HCMV depends on the host cell to provide macromolecular building blocks for virion production, and throughout the course of its evolution, HCMV has adapted to manipulate numerous fundamental cellular processes, including RNA accumulation (4), translation (5), metabolism (6, 7), secretory pathways (8), and the cell cycle (9).The nuclear receptor gene family, of which there are 48 members in the human genome, encodes transcription factors (10). Some bind to specific hydrophobic ligands such as steroids, vitamin D, fatty acids, and lipids, providing a means to sense changes in the extracellular or intracellular environment; others are "orphans" with no known ligand, and are often constitutively active (11). As transcription factors that modulate broad gene regulatory networks, nuclear receptors control numerous physiological processes, including aspects of metabolism (cell autonomously and at the whole-organism level), development, and cellular differentiation, cancer, circadian rhythm, and immunity (11).The first indication that nuclear receptor signaling modulates HCMV replication surfaced after the identification of a retinoic acid response element within the major immediate-early promoter (MIEP) that caused a dose-dependent response to retinoic acid in reporter assays (12). Retinoic acid was further shown to enhance viral DNA replication and progeny production in human embryonal carcinoma cells and foreskin fibroblasts (13,14). The ligand also exacerbated symptoms of infection,...

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Section: Methodsmentioning

confidence: 99%

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confidence: 99%

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confidence: 99%

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Estrogen-related receptor α is required for efficient human cytomegalovirus replication

Hwang¹,

Purdy

Wu³

et al. 2014

Proc. Natl. Acad. Sci. U.S.A.

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“…In larger well formats, reagents were scaled up based on growth area. Two siRNA libraries were used: Mission Metabolism and Cell Trafficking Panel and Mission Transferase Panel (Sigma) for a total of 401 cellular enzymes (7). Three days after transfection, cells were infected with HSV-1 at a multiplicity of 0.02 IU per cell.…”

Section: Discussionmentioning

confidence: 99%

Argininosuccinate synthetase 1 depletion produces a metabolic state conducive to herpes simplex virus 1 infection

Grady¹,

Purdy²,

Rabinowitz

et al. 2013

Proc. Natl. Acad. Sci. U.S.A.

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Herpes simplex virus 1 (HSV-1) infection triggers specific metabolic changes in its host cell. To explore the interactions between cellular metabolism and HSV-1 infection, we performed an siRNA screen of cellular metabolic genes, measuring their effect on viral replication. The screen identified multiple enzymes predicted to influence HSV-1 replication, including argininosuccinate synthetase 1 (AS1), which consumes aspartate as part of de novo arginine synthesis. Knockdown of AS1 robustly enhanced viral genome replication and the production of infectious virus. Using high-resolution liquid chromatography-mass spectrometry, we found that the metabolic phenotype induced by knockdown of AS1 in human fibroblasts mimicked multiple aspects of the metabolic program observed during HSV-1 infection, including an increase in multiple nucleotides and their precursors. Together with the observation that AS1 protein and mRNA levels decrease during wild-type infection, this work suggests that reduced AS1 activity is partially responsible for the metabolic program induced by infection.metabolomics | herpesviruses M any viruses, including herpes simplex virus 1 (HSV-1), human cytomegalovirus (HCMV), influenza, Dengue, hepatitis C, and hepatitis E (1-11), have been shown to significantly perturb metabolic homeostasis over the course of infection. HSV-1 is an intriguing case study, because the virus encodes several of its own metabolic enzymes, including a dUTPase, uracil-DNA glycosylase, ribonucleotide reductase, and thymidine kinase (12). Infection of host cells by multiple strains of HSV-1 induces alterations to central carbon metabolism, such as increased flux through upper glycolysis, and the anapleurotic feeding of carbons into the citric acid (TCA) cycle through the activity of pyruvate carboxylase (1). Infection also induces nucleotide synthesis, manifested by the increased flux of carbons through aspartate to nucleotides, as well as the increased pool levels of multiple nucleotides themselves (1).Although the metabolic reprogramming that occurs during HSV-1 infection has been described, the importance of specific host cell metabolic enzymes to infection is relatively unknown. Here we sought to further resolve the relationship between infection and metabolism by screening for cellular metabolic enzymes that modulate the efficiency of viral infection in primary human fibroblasts. The screen identified several dozen enzymes that modulate HSV-1 growth, including argininosuccinate synthetase (AS1), which antagonized viral replication.AS1 acts as a homotetramer to catalyze the synthesis of argininosuccinate from aspartate and citrulline (13,14). The enzyme plays a role in the production of urea in the kidney and liver, but functions primarily as the limiting step in the citrulline-nitric oxide (NO) cycle in other cell types (15). As a constituent of the citrulline-NO cycle, AS1 participates in the de novo production of the nonessential amino acid arginine as well as the soluble factor NO.We found that AS1 knockdown by siRNA in...

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“…Likewise, carbons from glutamine are redirected for anaplerotic usage to replenish the TCA cycle (Munger et al 2008;Chambers et al 2010). HCMV infection in human fibroblast dramatically increases the elongation of fatty acids to generate very long chain fatty acid tails (VLCFAs) (Koyuncu et al 2013;Purdy et al 2015). These VLCFAs are used to build the viral envelope.…”

Section: Manipulation Of Host Metabolismmentioning

confidence: 99%

Recent advances in CMV tropism, latency, and diagnosis during aging

et al. 2017

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Human cytomegalovirus (CMV) is one of the largest viruses known to cause human diseases. Chronic CMV infection, as defined by anti-CMV IgG serology, increases with age and is highly prevalent in older adults. It has complex biology with significant immunologic and health consequences. This article aims to summarize research findings presented at the 6th International Workshop on CMV and Immunosenescence that relate to advances in the areas of CMV tropism, latency, CMV manipulation of cell metabolism, and T cell memory inflation, as well as novel diagnostic evaluation and translational research of chronic CMV infection in older adults. Information summarized here represents the current state of knowledge in these important fields. Investigators have also identified a number of areas that deserve further and more in-depth investigation, including building more precise parallels between mouse CMV (mCMV) and human CMV (HCMV) research. It is hoped that this article will also stimulate engaging discussion on strategies and direction to advance the science to the next level.

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Saturated Very Long Chain Fatty Acids Are Required for the Production of Infectious Human Cytomegalovirus Progeny

Cited by 94 publications

References 60 publications

Estrogen-related receptor α is required for efficient human cytomegalovirus replication

Estrogen-related receptor α is required for efficient human cytomegalovirus replication

Argininosuccinate synthetase 1 depletion produces a metabolic state conducive to herpes simplex virus 1 infection

Recent advances in CMV tropism, latency, and diagnosis during aging

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