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Cited by 114Cytomegalovirus and herpesvirus researchCancer, Lipids, and MetabolismLipid metabolism and biosynthesis

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

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Koyuncu, Emre, Purdy, John G., Rabinowitz, Joshua D., Shenk, Thomas·2013·PLoS Pathogens
DOI 10.1371/journal.ppat.1003333PMC PMC3656100
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Abstract

Human cytomegalovirus hijacks host cell metabolism, increasing the flux of carbon from glucose to malonyl-CoA, the committed precursor to fatty acid synthesis and elongation. Inhibition of acetyl-CoA carboxylase blocks the production of progeny virus. To probe further the role of fatty acid metabolism during infection, we performed an siRNA screen to identify host cell metabolic enzymes needed for the production of infectious cytomegalovirus progeny. The screen predicted that multiple long chain acyl-CoA synthetases and fatty acid elongases are needed during infection, and the levels of RNAs encoding several of these enzymes were upregulated by the virus. Roles for acyl-CoA synthetases and elongases during infection were confirmed by using small molecule antagonists. Consistent with a role for these enzymes, mass spectrometry-based fatty acid analysis with ¹³C-labeling revealed that malonyl-CoA is consumed by elongases to produce very long chain fatty acids, generating an approximately 8-fold increase in C26-C34 fatty acid tails in infected cells. The virion envelope was yet further enriched in C26-C34 saturated fatty acids, and elongase inhibitors caused the production of virions with lower levels of these fatty acids and markedly reduced infectivity. These results reveal a dependence of cytomegalovirus on very long chain fatty acid metabolism.

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Research works printed but not linked: 57

  1. [1]Terry LJ, Vastag L, Rabinowitz JD, Shenk T (2012) Human kinome profiling identifies a requirement for AMP-activated protein kinase during human cytomegalovirus infection. Proc Natl Acad Sci U S A 109: 3071– 3076. 22315427 PMC3286917
  2. [2]McArdle J, Moorman NJ, Munger J (2012) HCMV targets the metabolic stress response through activation of AMPK whose activity is important for viral replication. PLoS Pathog 8: e1002502. 22291597 PMC3266935
  3. [3]Munger J, Bajad SU, Coller HA, Shenk T, Rabinowitz JD (2006) Dynamics of the cellular metabolome during human cytomegalovirus infection. PLoS Pathog 2: e132. 17173481 PMC1698944
  4. [4]Munger J, Bennett BD, Parikh A, Feng XJ, McArdle J, et al (2008) Systems-level metabolic flux profiling identifies fatty acid synthesis as a target for antiviral therapy. Nat Biotechnol 26: 1179– 1186. 18820684 PMC2825756
  5. [5]Vastag L, Koyuncu E, Grady SL, Shenk TE, Rabinowitz JD (2011) Divergent effects of human cytomegalovirus and herpes simplex virus-1 on cellular metabolism. PLoS Pathog 7: e1002124. 21779165 PMC3136460
  6. [6]Chambers JW, Maguire TG, Alwine JC (2010) Glutamine metabolism is essential for human cytomegalovirus infection. J Virol 84: 1867– 1873. 19939921 PMC2812398
  7. [7]Spencer CM, Schafer XL, Moorman NJ, Munger J (2011) Human cytomegalovirus induces the activity and expression of acetyl-coenzyme A carboxylase, a fatty acid biosynthetic enzyme whose inhibition attenuates viral replication. J Virol 85: 5814– 5824. 21471234 PMC3126312
  8. [8]Soupene E, Kuypers FA (2008) Mammalian long-chain acyl-CoA synthetases. Exp Biol Med (Maywood) 233: 507– 521. 18375835 PMC3377585
  9. [9]Kazantzis M, Stahl A (2012) Fatty acid transport proteins, implications in physiology and disease. Biochim Biophys Acta 1821: 852– 857. 21979150 PMC3274620
  10. [10]Jakobsson A, Westerberg R, Jacobsson A (2006) Fatty acid elongases in mammals: their regulation and roles in metabolism. Prog Lipid Res 45: 237– 249. 16564093
  11. [12]Takeuchi K, Reue K (2009) Biochemistry, physiology, and genetics of GPAT, AGPAT, and lipin enzymes in triglyceride synthesis. Am J Physiol Endocrinol Metab 296: E1195– 1209. 19336658 PMC2692402
  12. [13]Tvrdik P, Asadi A, Kozak LP, Nedergaard J, Cannon B, et al (1997) Cig30, a mouse member of a novel membrane protein gene family, is involved in the recruitment of brown adipose tissue. J Biol Chem 272: 31738– 31746. 9395518
  13. [14]Van Horn CG, Caviglia JM, Li LO, Wang S, Granger DA, et al (2005) Characterization of recombinant long-chain rat acyl-CoA synthetase isoforms 3 and 6: identification of a novel variant of isoform 6. Biochemistry 44: 1635– 1642. 15683247
  14. [15]Kim JH, Lewin TM, Coleman RA (2001) Expression and characterization of recombinant rat Acyl-CoA synthetases 1, 4, and 5. Selective inhibition by triacsin C and thiazolidinediones. J Biol Chem 276: 24667– 24673. 11319222
  15. [16]Igal RA, Wang P, Coleman RA (1997) Triacsin C blocks de novo synthesis of glycerolipids and cholesterol esters but not recycling of fatty acid into phospholipid: evidence for functionally separate pools of acyl-CoA. Biochem J 324 (Pt 2) 529– 534. 9182714 PMC1218462
  16. [17]Nagase T, Takahashi T, Sasaki T, Nagumo A, Shimamura K, et al (2009) Synthesis and biological evaluation of a novel 3-sulfonyl-8-azabicyclo[3.2.1]octane class of long chain fatty acid elongase 6 (ELOVL6) inhibitors. J Med Chem 52: 4111– 4114. 19522500
  17. [18]Mizutani T, Ishikawa S, Nagase T, Takahashi H, Fujimura T, et al (2009) Discovery of novel benzoxazinones as potent and orally active long chain fatty acid elongase 6 inhibitors. J Med Chem 52: 7289– 7300. 19883081
  18. [19]Takahashi T, Nagase T, Sasaki T, Nagumo A, Shimamura K, et al (2009) Synthesis and evaluation of a novel indoledione class of long chain fatty acid elongase 6 (ELOVL6) inhibitors. J Med Chem 52: 3142– 3145. 19388647
  19. [20]Kamphorst JJ, Fan J, Lu W, White E, Rabinowitz JD (2011) Liquid chromatography-high resolution mass spectrometry analysis of fatty acid metabolism. Anal Chem 83: 9114– 9122. 22004349 PMC3230881
  20. [21]Sanchez V, Greis KD, Sztul E, Britt WJ (2000) Accumulation of virion tegument and envelope proteins in a stable cytoplasmic compartment during human cytomegalovirus replication: characterization of a potential site of virus assembly. J Virol 74: 975– 986. 10623760 PMC111618
  21. [22]Krzyzaniak MA, Mach M, Britt WJ (2009) HCMV-encoded glycoprotein M (UL100) interacts with Rab11 effector protein FIP4. Traffic 10: 1439– 1457. 19761540 PMC4118585
  22. [23]Das S, Vasanji A, Pellett PE (2007) Three-dimensional structure of the human cytomegalovirus cytoplasmic virion assembly complex includes a reoriented secretory apparatus. J Virol 81: 11861– 11869. 17715239 PMC2168812
  23. [24]Krzyzaniak M, Mach M, Britt WJ (2007) The cytoplasmic tail of glycoprotein M (gpUL100) expresses trafficking signals required for human cytomegalovirus assembly and replication. J Virol 81: 10316– 10328. 17626081 PMC2045486
  24. [25]Walther TC, Farese Jr RV (2012) Lipid Droplets and Cellular Lipid Metabolism. Annu Rev Biochem 81: 687– 714. 22524315 PMC3767414
  25. [26]Samsa MM, Mondotte JA, Iglesias NG, Assuncao-Miranda I, Barbosa-Lima G, et al (2009) Dengue virus capsid protein usurps lipid droplets for viral particle formation. PLoS Pathog 5: e1000632. 19851456 PMC2760139
  26. [27]Ogawa K, Hishiki T, Shimizu Y, Funami K, Sugiyama K, et al (2009) Hepatitis C virus utilizes lipid droplet for production of infectious virus. Proc Jpn Acad Ser B Phys Biol Sci 85: 217– 228. PMC3561845 19644222
  27. [28]Yu Y, Maguire TG, Alwine JC (2012) Human cytomegalovirus infection induces adipocyte-like lipogenesis through activation of sterol regulatory element binding protein 1. J Virol 86: 2942– 2949. 22258239 PMC3302344
  28. [29]Koopman R, Schaart G, Hesselink MK (2001) Optimisation of oil red O staining permits combination with immunofluorescence and automated quantification of lipids. Histochem Cell Biol 116: 63– 68. 11479724
  29. [30]Supuran CT (2008) Carbonic anhydrases: novel therapeutic applications for inhibitors and activators. Nat Rev Drug Discov 7: 168– 181. 18167490
  30. [31]Cao J, Li JL, Li D, Tobin JF, Gimeno RE (2006) Molecular identification of microsomal acyl-CoA:glycerol-3-phosphate acyltransferase, a key enzyme in de novo triacylglycerol synthesis. Proc Natl Acad Sci U S A 103: 19695– 19700. 17170135 PMC1702318
  31. [32]Pei Z, Fraisl P, Berger J, Jia Z, Forss-Petter S, et al (2004) Mouse very long-chain Acyl-CoA synthetase 3/fatty acid transport protein 3 catalyzes fatty acid activation but not fatty acid transport in MA-10 cells. J Biol Chem 279: 54454– 54462. 15469937
  32. [33]Pei Z, Sun P, Huang P, Lal B, Laterra J, et al (2009) Acyl-CoA synthetase VL3 knockdown inhibits human glioma cell proliferation and tumorigenicity. Cancer Res 69: 9175– 9182. 19920185 PMC2795097
  33. [34]Cobbs CS (2011) Evolving evidence implicates cytomegalovirus as a promoter of malignant glioma pathogenesis. Herpesviridae 2: 10. 22030012 PMC3214144
  34. [35]Pizer ES, Thupari J, Han WF, Pinn ML, Chrest FJ, et al (2000) Malonyl-coenzyme-A is a potential mediator of cytotoxicity induced by fatty-acid synthase inhibition in human breast cancer cells and xenografts. Cancer Res 60: 213– 218. 10667561
  35. [38]Gaigg B, Toulmay A, Schneiter R (2006) Very long-chain fatty acid-containing lipids rather than sphingolipids per se are required for raft association and stable surface transport of newly synthesized plasma membrane ATPase in yeast. J Biol Chem 281: 34135– 34145. 16980694
  36. [39]Ibanez CE, Schrier R, Ghazal P, Wiley C, Nelson JA (1991) Human cytomegalovirus productively infects primary differentiated macrophages. J Virol 65: 6581– 6588. 1658363 PMC250717
  37. [40]Lathey JL, Spector SA (1991) Unrestricted replication of human cytomegalovirus in hydrocortisone-treated macrophages. J Virol 65: 6371– 6375. 1656103 PMC250364
  38. [41]Ecker J, Liebisch G, Englmaier M, Grandl M, Robenek H, et al (2010) Induction of fatty acid synthesis is a key requirement for phagocytic differentiation of human monocytes. Proc Natl Acad Sci U S A 107: 7817– 7822. 20385828 PMC2867858
  39. [42]Yu D, Smith GA, Enquist LW, Shenk T (2002) Construction of a self-excisable bacterial artificial chromosome containing the human cytomegalovirus genome and mutagenesis of the diploid TRL/IRL13 gene. J Virol 76: 2316– 2328. 11836410 PMC153828
  40. [44]Schroer J, Shenk T (2008) Inhibition of cyclooxygenase activity blocks cell-to-cell spread of human cytomegalovirus. Proc Natl Acad Sci U S A 105: 19468– 19473. 19033472 PMC2614784
  41. [45]Wiebusch L, Truss M, Hagemeier C (2004) Inhibition of human cytomegalovirus replication by small interfering RNAs. J Gen Virol 85: 179– 184. 14718633
  42. [46]Zhang JH, Chung TD, Oldenburg KR (1999) A Simple Statistical Parameter for Use in Evaluation and Validation of High Throughput Screening Assays. J Biomol Screen 4: 67– 73. 10838414
  43. [47]Birmingham A, Selfors LM, Forster T, Wrobel D, Kennedy CJ, et al (2009) Statistical methods for analysis of high-throughput RNA interference screens. Nat Methods 6: 569– 575. 19644458 PMC2789971
  44. [48]Arvidsson S, Kwasniewski M, Riano-Pachon DM, Mueller-Roeber B (2008) QuantPrime–a flexible tool for reliable high-throughput primer design for quantitative PCR. BMC Bioinformatics 9: 465. 18976492 PMC2612009
  45. [49]Womack A, Shenk T (2010) Human cytomegalovirus tegument protein pUL71 is required for efficient virion egress. MBio 1: e00282. 21151777 PMC2999941
  46. [50]Koyuncu OO, Dobner T (2009) Arginine methylation of human adenovirus type 5 L4 100-kilodalton protein is required for efficient virus production. J Virol 83: 4778– 4790. 19264777 PMC2682095
  47. [51]Zhu H, Shen Y, Shenk T (1995) Human cytomegalovirus IE1 and IE2 proteins block apoptosis. J Virol 69: 7960– 7970. 7494309 PMC189741
  48. [52]Nevels M, Paulus C, Shenk T (2004) Human cytomegalovirus immediate-early 1 protein facilitates viral replication by antagonizing histone deacetylation. Proc Natl Acad Sci U S A 101: 17234– 17239. 15572445 PMC535392
  49. [53]Nowak B, Sullivan C, Sarnow P, Thomas R, Bricout F, et al (1984) Characterization of monoclonal antibodies and polyclonal immune sera directed against human cytomegalovirus virion proteins. Virology 132: 325– 338. 6322416
  50. [54]Silva MC, Yu QC, Enquist L, Shenk T (2003) Human cytomegalovirus UL99-encoded pp28 is required for the cytoplasmic envelopment of tegument-associated capsids. J Virol 77: 10594– 10605. 12970444 PMC228509
  51. [55]Munger J, Yu D, Shenk T (2006) UL26-deficient human cytomegalovirus produces virions with hypophosphorylated pp28 tegument protein that is unstable within newly infected cells. J Virol 80: 3541– 3548. 16537622 PMC1440364
  52. [56]Liu ST, Sharon-Friling R, Ivanova P, Milne SB, Myers DS, et al (2011) Synaptic vesicle-like lipidome of human cytomegalovirus virions reveals a role for SNARE machinery in virion egress. Proc Natl Acad Sci U S A 108: 12869– 12874. 21768361 PMC3150939
  53. [57]Folch J, Lees M, Sloane Stanley GH (1957) A simple method for the isolation and purification of total lipides from animal tissues. J Biol Chem 226: 497– 509. 13428781
  54. [58]Lu W, Clasquin MF, Melamud E, Amador-Noguez D, Caudy AA, et al (2010) Metabolomic analysis via reversed-phase ion-pairing liquid chromatography coupled to a stand alone orbitrap mass spectrometer. Anal Chem 82: 3212– 3221. 20349993 PMC2863137
  55. [59]Melamud E, Vastag L, Rabinowitz JD (2010) Metabolomic analysis and visualization engine for LC-MS data. Anal Chem 82: 9818– 9826. 21049934 PMC5748896
  56. [60]Bartz R, Seemann J, Zehmer JK, Serrero G, Chapman KD, et al (2007) Evidence that mono-ADP-ribosylation of CtBP1/BARS regulates lipid storage. Mol Biol Cell 18: 3015– 3025. 17538025 PMC1949384
  57. [61]Britt WJ (1984) Neutralizing antibodies detect a disulfide-linked glycoprotein complex within the envelope of human cytomegalovirus. Virology 135: 369– 378. 6330979

Cited by

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Authors
Koyuncu, E., Purdy, J. G., Rabinowitz, J. D., & Shenk, T.
Affiliations
Princeton University (US)
Corresponding author
Shenk, Thomas
ORCID
Venue
PLoS Pathogens
Publisher
Public Library of Science (PLoS)
Published
2013-05-16
Volume
9
Issue
5
Pages
e1003333
ISSN
  • 1553-7366 (ISSN-L)
  • 1553-7374
DOI
10.1371/journal.ppat.1003333
PMC
PMC3656100
OpenAlex
W2086333472
Cited by
114
References
4
Citations by year
2026: 5 · 2025: 9 · 2024: 7 · 2023: 11 · 2022: 6 · 2021: 14 · 2020: 7 · 2019: 12 · 2018: 12 · 2017: 7 · 2016: 9 · 2015: 6 · 2014: 8 · 2013: 1
FWCI
5.84
Type
journal article
Language
en
Open access
Licence
Locations
  • PLoS Pathogens (publishedVersion, cc-by)
  • Europe PMC (PubMed Central) (publishedVersion, cc-by)
  • PubMed Central (publishedVersion, cc-by)
  • PubMed (publishedVersion)
  • DOAJ (DOAJ: Directory of Open Access Journals) (submittedVersion)
  • Figshare (submittedVersion, cc-by)
Funders
  • American Heart Association (12POST9190001)
  • National Institutes of Health
  • National Cancer Institute (CA 82396, R01CA082396)
  • National Institute of General Medical Sciences (P50GM071508, GM71508)
  • National Institute of Allergy and Infectious Diseases (R01AI097382, AI97382, AI78063, R01AI078063)
Topics
  • Health Sciences › Medicine › Epidemiology › Cytomegalovirus and herpesvirus research
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Cancer Research › Cancer, Lipids, and Metabolism
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Biochemistry › Lipid metabolism and biosynthesis
Keywords
human cytomegalovirus, very long chain fatty acids, fatty acid elongation, acyl-CoA synthetase, fatty acid elongase, malonyl-CoA, acetyl-CoA carboxylase, siRNA screen
MeSH terms
Cell Line, Coenzyme A Ligases, Cytomegalovirus Infections, Cytomegalovirus, Fatty Acids, Humans, Malonyl Coenzyme A, Virion, RNA, Small Interfering

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