Preferences help
enabled [disable] Abstract
Number of results
2007 | 54 | 4 | 813-819
Article title

Effect of N-glycosylation inhibition on the synthesis and processing of classical swine fever virus glycoproteins

Title variants
Languages of publication
Classical swine fever virus (CSFV) is often used as a surrogate model in molecular studies of the closely related hepatitis C virus. In this report we have examined the effect of the inhibition of glycosylation on the survival and maturation of CSFV. Viral glycoproteins (Erns, E1, E2) form biologically active complexes - homo- and heterodimers, which are indispensable for viral life cycle. Those complexes are highly N-glycosylated. We studied the influence of N-glycosylation on dimer formation using Erns and E2 glycoproteins produced in insect cells after infection with recombinant baculoviruses. The glycoproteins were efficiently synthesized in insect cells, had similar molecular masses and formed dimers like their natural counterparts. Surprisingly, the addition of tunicamycin (an antibiotic which blocks early steps of glycosylation) to insect cell culture blocked not only dimer formation but it also led to an almost complete disappearance of E2 even in monomeric form. Tunicamycin did not exert a similar effect on the synthesis and formation of Erns dimers; the dimers were still formed, which suggests that Erns glycan chains are not necessary for dimer formation. We have also found that very low doses of tunicamycin (much lower than those used for blocking N-glycosylation) drastically reduced CSFV spread in SK6 (swine kidney) cell culture and the virus yield. These facts indicate that N-glycosylation inhibitors structurally similar to tunicamycin may be potential therapeutics for the inhibition of the spread of CSFV and related viruses.
Physical description
  • Elbein AD (1987) Inhibitors of the biosynthesis and processing of N-linked oligosaccharide chains. Annu Rev Biochem 56: 497-534.
  • Frey CF, Bauhofer O, Ruggli N, Summerfield A, Hofmann MA, Tratschin JD (2006) Classical swine fever virus replicon particles lacking the E^rns gene: a potential marker vaccine for intradermal application. Vet Res 37: 655-670.
  • Horikawa K, Oishi N, Nakagawa J, Kasai A, Hayakawa K, Hiramatsu N, Takano Y, Yokouchi M, Yao J, Kitamura M (2006) Novel potential of tunicamycin as an activator of the aryl hydrocarbon receptor-dioxin responsive element signaling pathway. FEBS Lett 580: 3721-3725.
  • Hulst MM, Moormann RJ (1997) Inhibition of pestivirus infection in cell culture by envelope proteins E(rns) and E2 of classical swine fever virus E(rns) and E2 interact with different receptors. J Gen Virol 78: 2779-2787.
  • Hulst MM, Himes G, Newbigin E, Moormann RJ (1994) Glycoprotein E2 of classical swine fever virus: expression in cells and identification as a ribonuclease. Virology 200: 558-565.
  • Imperiali B, O'Connor SE, Hendrickson T, Kellenberger C (1999) Chemistry and biology of asparagine-linked glycosylation. Pure Appl Chem 71: 777-787.
  • Indyk K, Olczak T, Ciuraszkiewicz J, Watorek W, Olczak M (2007) Analysis of individual azurocidin N-glycosylation sites in regard to its secretion by insect cells, susceptibility to proteolysis and antibacterial activity. Acta Biochim Polon 54: 567-573.
  • Jones J, Krag S, Betenbaugh MJ (2005) Controlling N-linked glycan site occupancy. Biochim Biophys Acta 1726: 121-137.
  • Luckow VA, Lee SC, Barry GF, Olins PO (1993) Efficient generation of infectious recombinant baculoviruses by site-specific transposon-mediated insertion of foreign genes into a baculovirus genome propagated in Escherichia coli. J Virol 67: 4566-4579.
  • Leavitt R, Schlesinger S, Kornfeld S (1977) Tunicamycin inhibits glycosylation and multiplication of Sindibis and vesicular stomatitis viruses. J Virol 21: 375-385.
  • Maruo K, Nagata T, Yamamoto S, Nagai K, Yajima Y, Maruo S, Nishizaki T (2003) Tunicamycin inhibits NMDA and AMPA receptor responses independently of N-glycosylation. Brain Res 977: 294-297.
  • Mehta A, Zitzmann N, Rudd PM, Block TM, Dwek RA (1998) Alpha-glucosidase inhibitors as potential broad based anti-viral agents. FEBS Lett 430: 17-22.
  • Miller LK (1993) Baculoviruses: high level of expression in insect cells. Curr Opin Genet Dev 3: 97-101.
  • Pande AB, Carr V, Wong SA, Dalton K, Jones IM, McCauley JW Charleston B (2005) The glycosylation pattern of baculovirus expressed envelope protein E2 affects its ability to prevent infection with bovine viral diarrhoea virus. Virus Res 114: 54-62.
  • Parodi AJ (2000) Role of N-oligosaccharide endoplasmic reticulum processing reactions in glycoprotein folding and degradation. Biochem J 348: 1-13.
  • Risatti GR, Holinka LG, Fernandez Sainz I, Carrillo C, Lu Z, Borca MV (2007) N-linked glycosylation status of classical swine fever virus strain Brescia E2 glycoprotein influences virulence in swine. J Virol 81: 924-933.
  • Rumen T, Unger G, Strauss HJ, Thiel H (1993) Processing of the envelope glycoproteins of pestiviruses. J Virol 67: 3288-3294.
  • Scanlan CN, Offer J, Zitzmann N, Dwek RA (2007) Exploiting the defensive sugars of HIV-1 for drug and vaccine design. Nature 446: 1038-1045.
  • Schneider R, Unger G, Stark R, Schneider-Scherzer E, Thiel HJ (1993) Identification of a structural glycoprotein of an RNA virus as a ribonuclease. Science 261: 1169-1171.
  • Thiel HJ, Stark R, Weiland E, Rumenapf T, Meyers G (1991) Hog cholera virus: molecular composition of virions from a pestivirus. J Virol 65: 4705-4712.
  • Trombetta ES (2003) The contribution of N-glycans and their processing in the endoplasmatic reticulum to glycoprotein biosynthesis. Glycobiology 13: 77R-91R.
  • Tyborowska J, Bienkowska-Szewczyk K, Rychłowski M, van Oirschot JT, Rijsewijk FAM (2000) The extracellular part of glycoprotein E of bovine herpesvirus 1 is sufficient for complex formation with glycoprotein I but not for cell-to-cell spread. Arch Virol 145: 333-351.
  • Tyborowska J, Reszka N, Kochan G, Szewczyk B (2006) Formation of pseudorabies virus glycoprotein E/I complex in baculovirus recombinant system. Acta Virol 50: 169-174.
  • Wang Z, Nie Y, Wang P, Ding M, Deng H (2004) Characterization of classical swine fever virus entry by using pseudotyped viruses: E1 and E2 are sufficient to mediate viral entry. Virology 330: 332-341.
  • Windish JM, Schneider R, Stark R, Weiland E, Meyers G, Thiel HJ (1996) RNase of classical swine fever virus: biochemical characterization and inhibition by virus-neutralising antibodies. J Virol 70: 352-358.
  • Weiland E, Stark R, Haas B, Rumenapf T, Meyers G, Thiel HJ (1990) Pestivirus glycoprotein which induces neutralizing antibodies forms part of a disulfide-linked heterodimer. J Virol 64: 3563-3569.
  • Weiland F, Weiland E, Unger G, Saalmuller A, Thiel HJ. (1999) Localization of pestivirus envelope proteins E(rns) and E2 at the cell surface and on isolated particles. J Gen Virol 80: 1157-1165.
Document Type
Publication order reference
YADDA identifier
JavaScript is turned off in your web browser. Turn it on to take full advantage of this site, then refresh the page.