1887

Abstract

Recently the recruitment of lipid droplets (LDs) to sites of rotavirus (RV) replication was reported. LDs are polymorphic organelles that store triacylglycerols, cholesterol and cholesterol esters. The neutral fats are derived from palmitoyl-CoA, synthesized via the fatty acid biosynthetic pathway. RV-infected cells were treated with chemical inhibitors of the fatty acid biosynthetic pathway, and the effects on viral replication kinetics were assessed. Treatment with compound C75, an inhibitor of the fatty acid synthase enzyme complex (FASN), reduced RV infectivity 3.2-fold (  =  0.07) and modestly reduced viral RNA synthesis (1.2-fold). Acting earlier in the fatty acid synthesis pathway, TOFA [5-(Tetradecyloxy)-2-furoic acid] inhibits the enzyme acetyl-CoA carboxylase 1 (ACC1). TOFA reduced the infectivity of progeny RV 31-fold and viral RNA production 6-fold. The effect of TOFA on RV infectivity and RNA replication was dose-dependent, and infectivity was reduced by administering TOFA up to 4 h post-infection. Co-treatment of RV-infected cells with C75 and TOFA synergistically reduced viral infectivity. Knockdown by siRNA of FASN and ACC1 produced findings similar to those observed by inhibiting these proteins with the chemical compounds. Inhibition of fatty acid synthesis using a range of approaches uniformly had a more marked impact on viral infectivity than on viral RNA yield, inferring a role for LDs in virus assembly and/or egress. Specific inhibitors of fatty acid metabolism may help pinpoint the critical structural and biochemical features of LDs that are essential for RV replication, and facilitate the development of antiviral therapies.

Loading

Article metrics loading...

/content/journal/jgv/10.1099/vir.0.050146-0
2013-06-01
2019-10-16
Loading full text...

Full text loading...

/deliver/fulltext/jgv/94/6/1310.html?itemId=/content/journal/jgv/10.1099/vir.0.050146-0&mimeType=html&fmt=ahah

References

  1. Arnoldi F., Campagna M., Eichwald C., Desselberger U., Burrone O. R.. ( 2007;). Interaction of rotavirus polymerase VP1 with nonstructural protein NSP5 is stronger than that with NSP2. . J Virol 81:, 2128–2137. [CrossRef][PubMed]
    [Google Scholar]
  2. Barba G., Harper F., Harada T., Kohara M., Goulinet S., Matsuura Y., Eder G., Schaff Zs., Chapman M. J.. & other authors ( 1997;). Hepatitis C virus core protein shows a cytoplasmic localization and associates to cellular lipid storage droplets. . Proc Natl Acad Sci U S A 94:, 1200–1205. [CrossRef][PubMed]
    [Google Scholar]
  3. Bartenschlager R., Penin F., Lohmann V., André P.. ( 2011;). Assembly of infectious hepatitis C virus particles. . Trends Microbiol 19:, 95–103. [CrossRef][PubMed]
    [Google Scholar]
  4. Cheung W., Gill M., Esposito A., Kaminski C. F., Courousse N., Chwetzoff S., Trugnan G., Keshavan N., Lever A., Desselberger U.. ( 2010;). Rotaviruses associate with cellular lipid droplet components to replicate in viroplasms, and compounds disrupting or blocking lipid droplets inhibit viroplasm formation and viral replication. . J Virol 84:, 6782–6798. [CrossRef][PubMed]
    [Google Scholar]
  5. Chukkapalli V., Heaton N. S., Randall G.. ( 2012;). Lipids at the interface of virus-host interactions. . Curr Opin Microbiol 15:, 512–518. [CrossRef][PubMed]
    [Google Scholar]
  6. Clifford G. M., Londos C., Kraemer F. B., Vernon R. G., Yeaman S. J.. ( 2000;). Translocation of hormone-sensitive lipase and perilipin upon lipolytic stimulation of rat adipocytes. . J Biol Chem 275:, 5011–5015. [CrossRef][PubMed]
    [Google Scholar]
  7. Eichwald C., Vascotto F., Fabbretti E., Burrone O. R.. ( 2002;). Rotavirus NSP5: mapping phosphorylation sites and kinase activation and viroplasm localization domains. . J Virol 76:, 3461–3470. [CrossRef][PubMed]
    [Google Scholar]
  8. Eichwald C., Rodriguez J. F., Burrone O. R.. ( 2004;). Characterization of rotavirus NSP2/NSP5 interactions and the dynamics of viroplasm formation. . J Gen Virol 85:, 625–634. [CrossRef][PubMed]
    [Google Scholar]
  9. Estes M., Kapikian A.. ( 2007;). Rotaviruses. . In Fields Virology, , 5th edn., pp. 1917–1974. Edited by Knipe D. M., Lamb R. A., Howley P. M... Philadelphia:: Kluwer Health/Lippincott, Williams and Wilkins;.
    [Google Scholar]
  10. Flavin R., Peluso S., Nguyen P. L., Loda M.. ( 2010;). Fatty acid synthase as a potential therapeutic target in cancer. . Future Oncol 6:, 551–562. [CrossRef][PubMed]
    [Google Scholar]
  11. Goodman J. M.. ( 2009;). Demonstrated and inferred metabolism associated with cytosolic lipid droplets. . J Lipid Res 50:, 2148–2156. [CrossRef][PubMed]
    [Google Scholar]
  12. Guseva N. V., Rokhlin O. W., Glover R. A., Cohen M. B.. ( 2011;). TOFA (5-tetradecyl-oxy-2-furoic acid) reduces fatty acid synthesis, inhibits expression of AR, neuropilin-1 and Mcl-1 and kills prostate cancer cells independent of p53 status. . Cancer Biol Ther 12:, 80–85. [CrossRef][PubMed]
    [Google Scholar]
  13. Heaton N. S., Randall G.. ( 2011;). Multifaceted roles for lipids in viral infection. . Trends Microbiol 19:, 368–375. [CrossRef][PubMed]
    [Google Scholar]
  14. Kapadia S. B., Chisari F. V.. ( 2005;). Hepatitis C virus RNA replication is regulated by host geranylgeranylation and fatty acids. . Proc Natl Acad Sci U S A 102:, 2561–2566. [CrossRef][PubMed]
    [Google Scholar]
  15. Kibbe W. A.. ( 2007;). OligoCalc: an online oligonucleotide properties calculator. . Nucleic Acids Res 35: (Web Server issue), W43-W46. [CrossRef][PubMed]
    [Google Scholar]
  16. Krogerus C., Samuilova O., Pöyry T., Jokitalo E., Hyypiä T.. ( 2007;). Intracellular localization and effects of individually expressed human parechovirus 1 non-structural proteins. . J Gen Virol 88:, 831–841. [CrossRef][PubMed]
    [Google Scholar]
  17. Kuhajda F. P., Landree L. E., Ronnett G. V.. ( 2005;). The connections between C75 and obesity drug-target pathways. . Trends Pharmacol Sci 26:, 541–544. [CrossRef][PubMed]
    [Google Scholar]
  18. Martin S., Parton R. G.. ( 2006;). Lipid droplets: a unified view of a dynamic organelle. . Nat Rev Mol Cell Biol 7:, 373–378. [CrossRef][PubMed]
    [Google Scholar]
  19. Matthijnssens J., Otto P. H., Ciarlet M., Desselberger U., Van Ranst M., Johne R.. ( 2012;). VP6-sequence-based cutoff values as a criterion for rotavirus species demarcation. . Arch Virol 157:, 1177–1182. [CrossRef][PubMed]
    [Google Scholar]
  20. Miyanari Y., Atsuzawa K., Usuda N., Watashi K., Hishiki T., Zayas M., Bartenschlager R., Wakita T., Hijikata M., Shimotohno K.. ( 2007;). The lipid droplet is an important organelle for hepatitis C virus production. . Nat Cell Biol 9:, 1089–1097. [CrossRef][PubMed]
    [Google Scholar]
  21. Parashar U. D., Hummelman E. G., Bresee J. S., Miller M. A., Glass R. I.. ( 2003;). Global illness and deaths caused by rotavirus disease in children. . Emerg Infect Dis 9:, 565–572. [CrossRef][PubMed]
    [Google Scholar]
  22. Parker R. A., Kariya T., Grisar J. M., Petrow V.. ( 1977;). 5-(Tetradecyloxy)-2-furancarboxylic acid and related hypolipidemic fatty acid-like alkyloxyarylcarboxylic acids. . J Med Chem 20:, 781–791. [CrossRef][PubMed]
    [Google Scholar]
  23. Petrie B. L., Graham D. Y., Hanssen H., Estes M. K.. ( 1982;). Localization of rotavirus antigens in infected cells by ultrastructural immunocytochemistry. . J Gen Virol 63:, 457–467. [CrossRef][PubMed]
    [Google Scholar]
  24. Pizer E. S., Thupari J., Han W. F., Pinn M. L., Chrest F. J., Frehywot G. L., Townsend C. A., Kuhajda F. P.. ( 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.[PubMed]
    [Google Scholar]
  25. Reed L. J., Muench H.. ( 1938;). A simple method of estimating fifty per cent endpoints. . Am J Epidemiol 27:, 493–497.
    [Google Scholar]
  26. Ruiz-Palacios G. M., Pérez-Schael I., Velázquez F. R., Abate H., Breuer T., Clemens S. C., Cheuvart B., Espinoza F., Gillard P.. & other authors ( 2006;). Safety and efficacy of an attenuated vaccine against severe rotavirus gastroenteritis. . N Engl J Med 354:, 11–22. [CrossRef][PubMed]
    [Google Scholar]
  27. Samsa M. M., Mondotte J. A., Iglesias N. G., Assunção-Miranda I., Barbosa-Lima G., Da Poian A. T., Bozza P. T., Gamarnik A. V.. ( 2009;). Dengue virus capsid protein usurps lipid droplets for viral particle formation. . PLoS Pathog 5:, e1000632. [CrossRef][PubMed]
    [Google Scholar]
  28. Schmid B., Rippmann J. F., Tadayyon M., Hamilton B. S.. ( 2005;). Inhibition of fatty acid synthase prevents preadipocyte differentiation. . Biochem Biophys Res Commun 328:, 1073–1082. [CrossRef][PubMed]
    [Google Scholar]
  29. Schneider C. A., Rasband W. S., Eliceiri K. W.. ( 2012;). NIH Image to ImageJ: 25 years of image analysis. . Nat Methods 9:, 671–675. [CrossRef][PubMed]
    [Google Scholar]
  30. Simmonds P.. ( 2012;). SSE: a nucleotide and amino acid sequence analysis platform. . BMC Res Notes 5:, 50. [CrossRef][PubMed]
    [Google Scholar]
  31. Spencer C. M., Schafer X. L., Moorman N. J., 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. [CrossRef][PubMed]
    [Google Scholar]
  32. Tate J. E., Burton A. H., Boschi-Pinto C., Steele A. D., Duque J., Parashar U. D..WHO-coordinated Global Rotavirus Surveillance Network ( 2012;). 2008 estimate of worldwide rotavirus-associated mortality in children younger than 5 years before the introduction of universal rotavirus vaccination programmes: a systematic review and meta-analysis. . Lancet Infect Dis 12:, 136–141. [CrossRef][PubMed]
    [Google Scholar]
  33. Tennant J. R.. ( 1964;). Evaluation of the trypan blue technique for determination of cell viability. . Transplantation 2:, 685–694. [CrossRef][PubMed]
    [Google Scholar]
  34. Vesikari T., Matson D. O., Dennehy P., Van Damme P., Santosham M., Rodriguez Z., Dallas M. J., Heyse J. F., Goveia M. G.. & other authors ( 2006;). Safety and efficacy of a pentavalent human-bovine (WC3) reassortant rotavirus vaccine. . N Engl J Med 354:, 23–33. [CrossRef][PubMed]
    [Google Scholar]
  35. Yen C., Tate J. E., Patel M. M., Cortese M. M., Lopman B., Fleming J., Lewis K., Jiang B., Gentsch J.. & other authors ( 2011;). Rotavirus vaccines: update on global impact and future priorities. . Hum Vaccin 7:, 1282–1290. [CrossRef][PubMed]
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/jgv/10.1099/vir.0.050146-0
Loading
/content/journal/jgv/10.1099/vir.0.050146-0
Loading

Data & Media loading...

Supplements

Supplementary material 

PDF

Supplementary material 

PDF

Most Cited This Month

This is a required field
Please enter a valid email address
Approval was a Success
Invalid data
An Error Occurred
Approval was partially successful, following selected items could not be processed due to error