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Temperature-sensitive steps in the transport of Semliki Forest virus envelope proteins in mosquito C6/36 cells

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Summary

We have analysed the temperature dependence of the transport of Semliki Forest virus (SFV) envelope proteins in mosquito cells, the natural host cells of alphaviruses. These cells are cultivated at a lower temperature (28 °C) and have a different lipid composition as compared to mammalian cells. When the incubation temperature was reduced at early times after infection, the onset of virus shedding was delayed and the maximal titers decreased correspondingly to the temperature. No virus was shed at 12 °C. No evidence was observed for a block of virus release due to a shift of the sites of virus maturation. When the incubation temperature was reduced at later times after infection a critical temperature of 12 °C was again observed. At this temperature no transport of viral proteins took place, p62 remained uncleaved, the glycan processing of E1 did not occur and the envelope proteins accumulated in a pre-Golgi compartment. We suggest a mathematical formula which allows the extrapolation of transport data to the temperature at which intracellular protein transport becomes blocked.

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References

  1. Bonatti S, Migliaccio G, Simons K (1989) Palmitylation of viral membrane glycoproteins takes place after exit from the endoplasmic reticulum. J Biol Chem 264: 12590–12595

    PubMed  Google Scholar 

  2. Bordier C (1981) Phase separation of integral membrane proteins in Triton X-114 solution. J Biol Chem 256: 1604–1607

    PubMed  Google Scholar 

  3. Braakman I, Hoover-Litty H, Wagner KR, Helenius A (1991) Folding of influenza hemagglutinin in the endoplasmic reticulum. J Cell Biol 114: 401–411

    PubMed  Google Scholar 

  4. Ceriotti A, Colman A (1990) Trimer formation determines the rate of Influenza virus hemagglutinin transport in the early stages of secretion inXenopus oocytes. J Cell Biol 111: 409–420

    PubMed  Google Scholar 

  5. Chamberlain JP (1979) Fluorographic detection of radioactivity in polyacrylamide gels with the water soluble fluor, sodium salicylate. Anal Biochem 98: 132–135

    PubMed  Google Scholar 

  6. De Curtis I, Simons K (1988) Dissection of Semliki Forest virus glycoprotein delivery from the trans-Golgi network to the cell surface in permeabilized BHK cells. Proc Natl Acad Sci USA 85: 8052–8056

    PubMed  Google Scholar 

  7. Fast PG (1970) Insect lipids. Prog Chem Fats Lipids 11: 181–242

    Google Scholar 

  8. Fuller SD, Bravo R, Simons K (1985) An enzymatic assay reveals that proteins destined for apical or basolateral domains of an epithelial cell line share the same late Golgi compartments. EMBO J 42: 297–307

    Google Scholar 

  9. Garoff H, Simons K, Dobberstein B (1978) Assembly of the Semliki Forest virus membrane glycoproteins in the membrane of the endoplasmic reticulum in vitro. J Mol Biol 124: 587–600

    PubMed  Google Scholar 

  10. Gliedman JB, Smith JF, Brown DT (1975) Morphogenesis of Sindbis virus in culturedAedes albopictus cells. J Virol 16: 913–926

    PubMed  Google Scholar 

  11. Griffiths G, Pfeiffer S, Simons K, Matlin K (1985) Exit of newly synthesized membrane proteins from the trans cisterna of the Golgi complex to the plasma membrane. J Cell Biol 101: 949–964

    PubMed  Google Scholar 

  12. Igarashi A (1978) Isolation of a Singh'sAedes albopictus cell clone sensitive to Dengue and Chikungunya viruses. J Gen Virol 40: 531–544

    PubMed  Google Scholar 

  13. Koblet H, Kempf C, Kohler U, Omar A (1985) Conformational changes at pH 6 on the cell surface of Semliki Forest virus infectedAedes albopictus cells. Virology 143: 334–336

    PubMed  Google Scholar 

  14. Koblet H, Omar A, Kempf C (1987) Fusion of alphavirus-infected mosquito cells. In: Yunker CE (ed) Arboviruses in arthropod cells in vitro, vol 2. CRC Press, Boca Raton, pp 70–90

    Google Scholar 

  15. Koblet H (1990) The “merry-go-round”: alphaviruses between vertebrate and invertebrate cells. Adv Virus Res 38: 343–402

    PubMed  Google Scholar 

  16. Koblet H, Schärer CG, Vallan C, Naim HY (1992) Exocytotic transport of Semliki Forest virus (SFV) envelope proteins in mosquito (C6/36) cells. In: Fraser MJ Jr (ed) Proceedings of the 8th International Conference on Invertebrate and Fish Tissue Culture. Tissue Culture Association, Columbia, pp 195–203

    Google Scholar 

  17. Kornfeld R, Kornfeld S (1985) Assembly of asparagine-linked oligosaccharides. Annu Rev Biochem 54: 631–664

    PubMed  Google Scholar 

  18. Kuismanen E, Saraste J (1989) Low temperature-induced transport blocks as tools to manipulate membrane traffic. In: Tartakoff AM (ed) Methods in cell biology, vol 32. Academic Press, London, pp 257–274

    Google Scholar 

  19. Laemmli UL (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227: 680–685

    PubMed  Google Scholar 

  20. Liljeström P, Garoff H (1991) Internally located cleavable signal sequences direct the formation of Semliki Forest virus membrane proteins from a polyprotein precursor. J Virol 65: 147–154

    PubMed  Google Scholar 

  21. Lobigs M, Zhao H, Garoff H (1990) Function of Semliki Forest virus E3 peptide in virus assembly: replacement of E3 with an artificial signal peptide abolishes spike heterodimerization and surface expression of E1. J Virol 64: 4346–4355

    PubMed  Google Scholar 

  22. Luukkonen A, Kääriäinen L, Renkonen O (1976) Phospholipids of Semliki Forest virus grown in cultured mosquito cells. Biochim Biophys Acta 450: 109–120

    PubMed  Google Scholar 

  23. Matlin KS, Simons K (1983) Reduced temperature prevents transfer of a membrane glycoprotein to the cell surface but does not prevent terminal glycosylation. Cell 34: 233–243

    PubMed  Google Scholar 

  24. Miller ML, Brown DT (1992) Morphogenesis of Sindbis virus in three subclones ofAedes albopictus (mosquito) cells. J Virol 66: 4180–4190

    PubMed  Google Scholar 

  25. Moreau P, Juguelin H, Cassagne C, Morré DJ (1992) Molecular basis for low temperature compartment formation by transitional endoplasmic reticulum of rat liver. FEBS Lett 310: 223–228

    PubMed  Google Scholar 

  26. Mottet G, Tuffereau C, Roux L (1986) Reduced temperature can block different glycoproteins at different steps during transport to the plasma membrane. J Gen Virol 67: 2029–2035

    PubMed  Google Scholar 

  27. Naim HY, Sterchi E, Lentze M (1988) Biosynthesis of the human ucrase-isomaltase complex. J Biol Chem 263: 7242–7253

    PubMed  Google Scholar 

  28. Naim HY, Koblet H (1990) The cleavage of p62, the precursor of E2 and E3, is an early and continuous event in Semliki Forest virus-infectedAedes albopictus cells. Arch Virol 110: 221–237

    PubMed  Google Scholar 

  29. Naim HY, Koblet H (1992) Asparagine-linked oligosaccharides of Semliki Forest virus grown in mosquito cells. Arch Virol 122: 45–60

    PubMed  Google Scholar 

  30. Presley JF, Brown DT (1989) The proteolytic cleavage of PE2 to envelope glycoprotein E2 is not strictly required for the maturation of Sindbis virus. J Virol 63: 1975–1980

    PubMed  Google Scholar 

  31. Presley JF, Polo JM, Johnston RE, Brown DT (1991) Proteolytic processing of the Sindbis virus membrane protein precursor PE2 is nonessential for growth in vertebrate cells but is required for efficient growth in invertebrate cells. J Virol 65: 1905–1909

    PubMed  Google Scholar 

  32. Raghow RS, Grace TDC, Filshie BK, Bartley W, Dalgarno L (1973) Ross River virus replication in cultured mosquito and mammalian cells: virus growth and correlated ultrastructural changes. J Gen Virol 21: 109–122

    PubMed  Google Scholar 

  33. Rotundo RL, Fambrough DM (1980) Secretion of acetylcholinesterase: relation to acetylcholine receptor metabolism. Cell 22: 595–602

    PubMed  Google Scholar 

  34. Russel DL, Dalrymple JM, Johnson RE (1989) Sindbis virus mutations which coordinately affect glycoprotein processing, penetration and virulence in mice. J Virol 63: 1619–1629

    PubMed  Google Scholar 

  35. Saraste J, Kuismanen E (1984) Pre- and post-Golgi vacuoles operate in the transport of Semliki Forest virus membrane glycoproteins to the cell surface. Cell 38: 535–549

    PubMed  Google Scholar 

  36. Saraste J, Palade GE, Farquhar MG (1986) Temperature-sensitive steps in the transport of secretory proteins through the Golgi complex in exocrine pancreatic cells. Proc Natl Acad Sci USA 83: 6425–6429

    PubMed  Google Scholar 

  37. Sarver N, Stollar V (1977) Sindbis virus-induced cytopathic effect in clones ofAedes albopictus (Singh) cells. Virology 80: 390–400

    PubMed  Google Scholar 

  38. Schärer CG, Naim HY, Koblet H (1993) Palmitoylation of Semliki Forest virus glycoproteins in insect cells (C6/36) occurs in an early compartment and is coupled to the cleavage of the precursor p62. Arch Virol 132: 237–254

    PubMed  Google Scholar 

  39. Simizu B, Maeda S (1981) Growth patterns of temperature- sensitive mutants of western equine encephalitis virus in culturedAedes albopictus (mosquito) cells. J Gen Virol 56: 349–361

    PubMed  Google Scholar 

  40. Turell MJ (1989) Effect of environmental temperature on the vector competence ofAedes fowleri for Rift Valley Fever virus. Res Virol 140: 147–154

    PubMed  Google Scholar 

  41. Turell MJ, Rossi CA, Bailey CL (1985) Effect of extrinsic incubation temperature on the ability ofAedes taeniorhynchus andCulex pipiens to transmit Rift Valley Fever virus. Am J Trop Med Hyg 34: 1211–1218

    PubMed  Google Scholar 

  42. van't Hof W, van Meer G (1990) Generation of lipid polarity in intestinal epithelial (Caco−2) cells: sphingolipid synthesis in the Golgi complex and sorting before vesicular traffic to the plasma membrane. J Cell Biol 111: 977–986

    PubMed  Google Scholar 

  43. Wessel D, Flügge UI (1984) A method for the quantitative recovery of protein in dilute solution in the presence of detergents and lipids. Anal Biochem 138: 141–143

    PubMed  Google Scholar 

  44. Zhao H, Garoff H (1992) Role of cell surface spikes in alphavirus budding. J Virol 66: 7089–7095

    PubMed  Google Scholar 

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Author notes

  1. C. Vallan

    Present address: Institute of Anatomy, University of Berne, Switzerland

Authors and Affiliations

  1. Institute of Medical Microbiology, University of Berne, Berne, Switzerland

    C. Vallan, C. G. Schärer & H. Koblet

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  2. C. G. Schärer
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  3. H. Koblet
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Vallan, C., Schärer, C.G. & Koblet, H. Temperature-sensitive steps in the transport of Semliki Forest virus envelope proteins in mosquito C6/36 cells. Archives of Virology 134, 109–127 (1994). https://doi.org/10.1007/BF01379111

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  • DOI: https://doi.org/10.1007/BF01379111

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