Skip to main content
NCBI home page
Journal of Virology logoLink to Journal of Virology
. 1990 Aug;64(8):3643–3653. doi: 10.1128/jvi.64.8.3643-3653.1990

A conformational change in Sindbis virus glycoproteins E1 and E2 is detected at the plasma membrane as a consequence of early virus-cell interaction.

D C Flynn 1, W J Meyer 1, J M Mackenzie Jr 1, R E Johnston 1
PMCID: PMC249657  PMID: 1695253

Abstract

A conformational change in the structure of Sindbis (SB) virus was detected after virion attachment to baby hamster kidney cells but before internalization. The alteration was manifested as increased virion binding of certain glycoprotein E1 and E2 monoclonal antibodies (MAbs) that recognized transitional epitopes. These epitopes were inaccessible to MAb on native virions but became accessible to their cognate MAbs in the early stages of infection. Transit of virions through a low-pH compartment apparently was not required for the conformational change. Exposure of transitional epitopes was unaffected by treatment of BHK cells with NH4Cl and occurred normally in Chinese hamster ovary cells temperature sensitive for endosomal acidification. However, the rearrangement was correlated with both the time course and temperature dependence of SB virus penetration, and the rearrangement occurred earlier with an SB virus mutant having an accelerated penetration phenotype. In addition, MAb to a transitional epitope, a probe specific for rearranged particles, retarded penetration of infectious virions. These results suggested that the SB virus E1/E2 glycoprotein spike undergoes a structural rearrangement as a consequence of virion interaction with the cell surface and that this altered virion form may be an important early intermediate in an entry pathway leading to productive infection.

Full text

PDF
3643

Images in this article

3647Image
on p.3647

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Acheson N. H., Tamm I. Replication of Semliki Forest virus: an electron microscopic study. Virology. 1967 May;32(1):128–143. doi: 10.1016/0042-6822(67)90261-9. [DOI] [PubMed] [Google Scholar]
  2. Barbey-Morel C. L., Oeltmann T. N., Edwards K. M., Wright P. F. Role of respiratory tract proteases in infectivity of influenza A virus. J Infect Dis. 1987 Apr;155(4):667–672. doi: 10.1093/infdis/155.4.667. [DOI] [PubMed] [Google Scholar]
  3. Baric R. S., Trent D. W., Johnston R. E. A Sindbis virus variant with a cell-determined latent period. Virology. 1981 Apr 15;110(1):237–242. doi: 10.1016/0042-6822(81)90029-5. [DOI] [PubMed] [Google Scholar]
  4. Birrell G. B., Hedberg K. K., Griffith O. H. Pitfalls of immunogold labeling: analysis by light microscopy, transmission electron microscopy, and photoelectron microscopy. J Histochem Cytochem. 1987 Aug;35(8):843–853. doi: 10.1177/35.8.2439584. [DOI] [PubMed] [Google Scholar]
  5. Bodkin D. K., Nibert M. L., Fields B. N. Proteolytic digestion of reovirus in the intestinal lumens of neonatal mice. J Virol. 1989 Nov;63(11):4676–4681. doi: 10.1128/jvi.63.11.4676-4681.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Brown D. T., Waite M. R., Pfefferkorn E. R. Morphology and morphogenesis of Sindbis virus as seen with freeze-etching techniques. J Virol. 1972 Sep;10(3):524–536. doi: 10.1128/jvi.10.3.524-536.1972. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Bächi T., Gerhard W., Yewdell J. W. Monoclonal antibodies detect different forms of influenza virus hemagglutinin during viral penetration and biosynthesis. J Virol. 1985 Aug;55(2):307–313. doi: 10.1128/jvi.55.2.307-313.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Cassell S., Edwards J., Brown D. T. Effects of lysosomotropic weak bases on infection of BHK-21 cells by Sindbis virus. J Virol. 1984 Dec;52(3):857–864. doi: 10.1128/jvi.52.3.857-864.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Chanas A. C., Gould E. A., Clegg J. C., Varma M. G. Monoclonal antibodies to Sindbis virus glycoprotein E1 can neutralize, enhance infectivity, and independently inhibit haemagglutination or haemolysis. J Gen Virol. 1982 Jan;58(Pt 1):37–46. doi: 10.1099/0022-1317-58-1-37. [DOI] [PubMed] [Google Scholar]
  10. Chow M., Newman J. F., Filman D., Hogle J. M., Rowlands D. J., Brown F. Myristylation of picornavirus capsid protein VP4 and its structural significance. Nature. 1987 Jun 11;327(6122):482–486. doi: 10.1038/327482a0. [DOI] [PubMed] [Google Scholar]
  11. Coombs K., Mann E., Edwards J., Brown D. T. Effects of chloroquine and cytochalasin B on the infection of cells by Sindbis virus and vesicular stomatitis virus. J Virol. 1981 Mar;37(3):1060–1065. doi: 10.1128/jvi.37.3.1060-1065.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Crowell R. L., Philipson L. Specific alterations of coxsackievirus B3 eluted from HeLa cells. J Virol. 1971 Oct;8(4):509–515. doi: 10.1128/jvi.8.4.509-515.1971. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Davis N. L., Fuller F. J., Dougherty W. G., Olmsted R. A., Johnston R. E. A single nucleotide change in the E2 glycoprotein gene of Sindbis virus affects penetration rate in cell culture and virulence in neonatal mice. Proc Natl Acad Sci U S A. 1986 Sep;83(18):6771–6775. doi: 10.1073/pnas.83.18.6771. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. De Sena J., Mandel B. Studies on the in vitro uncoating of poliovirus. I. Characterization of the modifying factor and the modifying reaction. Virology. 1976 Apr;70(2):470–483. doi: 10.1016/0042-6822(76)90288-9. [DOI] [PubMed] [Google Scholar]
  15. Dougherty W. G., Willis L., Johnston R. E. Topographic analysis of tobacco etch virus capsid protein epitopes. Virology. 1985 Jul 15;144(1):66–72. doi: 10.1016/0042-6822(85)90305-8. [DOI] [PubMed] [Google Scholar]
  16. Edwards J., Mann E., Brown D. T. Conformational changes in Sindbis virus envelope proteins accompanying exposure to low pH. J Virol. 1983 Mar;45(3):1090–1097. doi: 10.1128/jvi.45.3.1090-1097.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Ey P. L., Prowse S. J., Jenkin C. R. Isolation of pure IgG1, IgG2a and IgG2b immunoglobulins from mouse serum using protein A-sepharose. Immunochemistry. 1978 Jul;15(7):429–436. doi: 10.1016/0161-5890(78)90070-6. [DOI] [PubMed] [Google Scholar]
  18. Fricks C. E., Hogle J. M. Cell-induced conformational change in poliovirus: externalization of the amino terminus of VP1 is responsible for liposome binding. J Virol. 1990 May;64(5):1934–1945. doi: 10.1128/jvi.64.5.1934-1945.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Fuller S. D. The T=4 envelope of Sindbis virus is organized by interactions with a complementary T=3 capsid. Cell. 1987 Mar 27;48(6):923–934. doi: 10.1016/0092-8674(87)90701-x. [DOI] [PubMed] [Google Scholar]
  20. Furlong D. B., Nibert M. L., Fields B. N. Sigma 1 protein of mammalian reoviruses extends from the surfaces of viral particles. J Virol. 1988 Jan;62(1):246–256. doi: 10.1128/jvi.62.1.246-256.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Gidwitz S., Polo J. M., Davis N. L., Johnston R. E. Differences in virion stability among Sindbis virus pathogenesis mutants. Virus Res. 1988 May;10(2-3):225–239. doi: 10.1016/0168-1702(88)90018-4. [DOI] [PubMed] [Google Scholar]
  22. Goding J. W. Conjugation of antibodies with fluorochromes: modifications to the standard methods. J Immunol Methods. 1976;13(3-4):215–226. doi: 10.1016/0022-1759(76)90068-5. [DOI] [PubMed] [Google Scholar]
  23. Guttman N., Baltimore D. A plasma membrane component able to bind and alter virions of poliovirus type 1: studies on cell-free alteration using a simplified assay. Virology. 1977 Oct 1;82(1):25–36. doi: 10.1016/0042-6822(77)90029-0. [DOI] [PubMed] [Google Scholar]
  24. Helenius A., Kartenbeck J., Simons K., Fries E. On the entry of Semliki forest virus into BHK-21 cells. J Cell Biol. 1980 Feb;84(2):404–420. doi: 10.1083/jcb.84.2.404. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Helenius A., Marsh M., White J. Inhibition of Semliki forest virus penetration by lysosomotropic weak bases. J Gen Virol. 1982 Jan;58(Pt 1):47–61. doi: 10.1099/0022-1317-58-1-47. [DOI] [PubMed] [Google Scholar]
  26. Hogle J. M., Chow M., Filman D. J. Three-dimensional structure of poliovirus at 2.9 A resolution. Science. 1985 Sep 27;229(4720):1358–1365. doi: 10.1126/science.2994218. [DOI] [PubMed] [Google Scholar]
  27. Kielian M. C., Keränen S., Käriäinen L., Helenius A. Membrane fusion mutants of Semliki Forest virus. J Cell Biol. 1984 Jan;98(1):139–145. doi: 10.1083/jcb.98.1.139. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Kielian M. C., Marsh M., Helenius A. Kinetics of endosome acidification detected by mutant and wild-type Semliki Forest virus. EMBO J. 1986 Dec 1;5(12):3103–3109. doi: 10.1002/j.1460-2075.1986.tb04616.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Kielian M., Helenius A. pH-induced alterations in the fusogenic spike protein of Semliki Forest virus. J Cell Biol. 1985 Dec;101(6):2284–2291. doi: 10.1083/jcb.101.6.2284. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Lonberg-Holm K., Gosser L. B., Kauer J. C. Early alteration of poliovirus in infected cells and its specific inhibition. J Gen Virol. 1975 Jun;27(3):329–342. doi: 10.1099/0022-1317-27-3-329. [DOI] [PubMed] [Google Scholar]
  31. Lonberg-Holm K., Korant B. D. Early interaction of rhinoviruses with host cells. J Virol. 1972 Jan;9(1):29–40. doi: 10.1128/jvi.9.1.29-40.1972. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Luukkonen A., von Bonsdorff C. H., Renkonen O. Characterization of Semliki Forest virus grown in mosquito cells. Comparison with the virus from hamster cells. Virology. 1977 May 1;78(1):331–335. doi: 10.1016/0042-6822(77)90105-2. [DOI] [PubMed] [Google Scholar]
  33. Marsh M., Bolzau E., Helenius A. Penetration of Semliki Forest virus from acidic prelysosomal vacuoles. Cell. 1983 Mar;32(3):931–940. doi: 10.1016/0092-8674(83)90078-8. [DOI] [PubMed] [Google Scholar]
  34. Müller G., Baigent C. L. Antigen controlled immuno diagnosis-- 'ACID test'. J Immunol Methods. 1980;37(2):185–190. doi: 10.1016/0022-1759(80)90203-3. [DOI] [PubMed] [Google Scholar]
  35. Ohkuma S., Poole B. Fluorescence probe measurement of the intralysosomal pH in living cells and the perturbation of pH by various agents. Proc Natl Acad Sci U S A. 1978 Jul;75(7):3327–3331. doi: 10.1073/pnas.75.7.3327. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Olmsted R. A., Baric R. S., Sawyer B. A., Johnston R. E. Sindbis virus mutants selected for rapid growth in cell culture display attenuated virulence in animals. Science. 1984 Jul 27;225(4660):424–427. doi: 10.1126/science.6204381. [DOI] [PubMed] [Google Scholar]
  37. Olmsted R. A., Meyer W. J., Johnston R. E. Characterization of Sindbis virus epitopes important for penetration in cell culture and pathogenesis in animals. Virology. 1986 Jan 30;148(2):245–254. doi: 10.1016/0042-6822(86)90322-3. [DOI] [PubMed] [Google Scholar]
  38. Polo J. M., Davis N. L., Rice C. M., Huang H. V., Johnston R. E. Molecular analysis of Sindbis virus pathogenesis in neonatal mice by using virus recombinants constructed in vitro. J Virol. 1988 Jun;62(6):2124–2133. doi: 10.1128/jvi.62.6.2124-2133.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  39. Rice C. M., Strauss J. H. Association of sindbis virion glycoproteins and their precursors. J Mol Biol. 1982 Jan 15;154(2):325–348. doi: 10.1016/0022-2836(82)90067-5. [DOI] [PubMed] [Google Scholar]
  40. Roehrig J. T., Gorski D., Schlesinger M. J. Properties of monoclonal antibodies directed against the glycoproteins of Sindbis virus. J Gen Virol. 1982 Apr;59(Pt 2):421–425. doi: 10.1099/0022-1317-59-2-421. [DOI] [PubMed] [Google Scholar]
  41. Roff C. F., Fuchs R., Mellman I., Robbins A. R. Chinese hamster ovary cell mutants with temperature-sensitive defects in endocytosis. I. Loss of function on shifting to the nonpermissive temperature. J Cell Biol. 1986 Dec;103(6 Pt 1):2283–2297. doi: 10.1083/jcb.103.6.2283. [DOI] [PMC free article] [PubMed] [Google Scholar]
  42. Rossmann M. G., Arnold E., Erickson J. W., Frankenberger E. A., Griffith J. P., Hecht H. J., Johnson J. E., Kamer G., Luo M., Mosser A. G. Structure of a human common cold virus and functional relationship to other picornaviruses. Nature. 1985 Sep 12;317(6033):145–153. doi: 10.1038/317145a0. [DOI] [PubMed] [Google Scholar]
  43. Schlesinger M. J., Schlesinger S., Burge B. W. Identification of a second glycoprotein in Sindbis virus. Virology. 1972 Feb;47(2):539–541. doi: 10.1016/0042-6822(72)90298-x. [DOI] [PubMed] [Google Scholar]
  44. Schmaljohn A. L., Johnson E. D., Dalrymple J. M., Cole G. A. Non-neutralizing monoclonal antibodies can prevent lethal alphavirus encephalitis. Nature. 1982 May 6;297(5861):70–72. doi: 10.1038/297070a0. [DOI] [PubMed] [Google Scholar]
  45. Schmaljohn A. L., Kokubun K. M., Cole G. A. Protective monoclonal antibodies define maturational and pH-dependent antigenic changes in Sindbis virus E1 glycoprotein. Virology. 1983 Oct 15;130(1):144–154. doi: 10.1016/0042-6822(83)90124-1. [DOI] [PubMed] [Google Scholar]
  46. Simmons D. T., Strauss J. H. Replication of Sindbis virus. I. Relative size and genetic content of 26 s and 49 s RNA. J Mol Biol. 1972 Nov 28;71(3):599–613. [PubMed] [Google Scholar]
  47. Skehel J. J., Bayley P. M., Brown E. B., Martin S. R., Waterfield M. D., White J. M., Wilson I. A., Wiley D. C. Changes in the conformation of influenza virus hemagglutinin at the pH optimum of virus-mediated membrane fusion. Proc Natl Acad Sci U S A. 1982 Feb;79(4):968–972. doi: 10.1073/pnas.79.4.968. [DOI] [PMC free article] [PubMed] [Google Scholar]
  48. Slade D. E., Johnston R. E., Dougherty W. G. Generation and characterization of monoclonal antibodies reactive with the 49-kDa proteinase of tobacco etch virus. Virology. 1989 Dec;173(2):499–508. doi: 10.1016/0042-6822(89)90562-x. [DOI] [PubMed] [Google Scholar]
  49. Stanley J., Cooper S. J., Griffin D. E. Alphavirus neurovirulence: monoclonal antibodies discriminating wild-type from neuroadapted Sindbis virus. J Virol. 1985 Oct;56(1):110–119. doi: 10.1128/jvi.56.1.110-119.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  50. Strauss E. G., Rice C. M., Strauss J. H. Complete nucleotide sequence of the genomic RNA of Sindbis virus. Virology. 1984 Feb;133(1):92–110. doi: 10.1016/0042-6822(84)90428-8. [DOI] [PubMed] [Google Scholar]
  51. Strauss J. H., Jr, Burge B. W., Pfefferkorn E. R., Darnell J. E., Jr Identification of the membrane protein and "core" protein of Sindbis virus. Proc Natl Acad Sci U S A. 1968 Feb;59(2):533–537. doi: 10.1073/pnas.59.2.533. [DOI] [PMC free article] [PubMed] [Google Scholar]
  52. Tycko B., Maxfield F. R. Rapid acidification of endocytic vesicles containing alpha 2-macroglobulin. Cell. 1982 Mar;28(3):643–651. doi: 10.1016/0092-8674(82)90219-7. [DOI] [PubMed] [Google Scholar]
  53. Webster R. G., Brown L. E., Jackson D. C. Changes in the antigenicity of the hemagglutinin molecule of H3 influenza virus at acidic pH. Virology. 1983 Apr 30;126(2):587–599. doi: 10.1016/s0042-6822(83)80015-4. [DOI] [PubMed] [Google Scholar]
  54. White J., Helenius A. pH-dependent fusion between the Semliki Forest virus membrane and liposomes. Proc Natl Acad Sci U S A. 1980 Jun;77(6):3273–3277. doi: 10.1073/pnas.77.6.3273. [DOI] [PMC free article] [PubMed] [Google Scholar]
  55. White J., Matlin K., Helenius A. Cell fusion by Semliki Forest, influenza, and vesicular stomatitis viruses. J Cell Biol. 1981 Jun;89(3):674–679. doi: 10.1083/jcb.89.3.674. [DOI] [PMC free article] [PubMed] [Google Scholar]
  56. Wirth D. F., Katz F., Small B., Lodish H. F. How a single Sindbis virus mRNA directs the synthesis of one soluble protein and two integral membrane glycoproteins. Cell. 1977 Feb;10(2):253–263. doi: 10.1016/0092-8674(77)90219-7. [DOI] [PubMed] [Google Scholar]
  57. Yewdell J. W., Gerhard W., Bachi T. Monoclonal anti-hemagglutinin antibodies detect irreversible antigenic alterations that coincide with the acid activation of influenza virus A/PR/834-mediated hemolysis. J Virol. 1983 Oct;48(1):239–248. doi: 10.1128/jvi.48.1.239-248.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  58. von Bonsdorff C. H., Harrison S. C. Sindbis virus glycoproteins form a regular icosahedral surface lattice. J Virol. 1975 Jul;16(1):141–145. doi: 10.1128/jvi.16.1.141-145.1975. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Journal of Virology are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES