Skip to main content
PMC home page
Journal of Virology logoLink to Journal of Virology
. 1991 Sep;65(9):4654–4664. doi: 10.1128/jvi.65.9.4654-4664.1991

Identification of antigenically important domains in the glycoproteins of Sindbis virus by analysis of antibody escape variants.

E G Strauss 1, D S Stec 1, A L Schmaljohn 1, J H Strauss 1
PMCID: PMC248920  PMID: 1714515

Abstract

To study important epitopes on glycoprotein E2 of Sindbis virus, eight variants selected to be singly or multiply resistant to six neutralizing monoclonal antibodies reactive against E2, as well as four revertants which had regained sensitivity to neutralization, were sequenced throughout the E2 region. To study antigenic determinants in glycoprotein E1, four variants selected for resistance to a neutralizing monoclonal antibody reactive with E1 were sequenced throughout the E2 and E1 regions. All of the salient changes in E2 occurred within a relatively small region between amino acids 181 and 216, a domain that encompasses a glycosylation site at residue 196 and that is rich in charged amino acids. Almost all variants had a change in charge, suggesting that the charged nature of this domain is important for interaction with antibodies. Variants independently isolated for resistance to the same antibody were usually altered in the same amino acid, and reversion to sensitivity occurred at the sites of the original mutations, but did not always restore the parental amino acid. The characteristics of this region suggest that this domain is found on the surface of E2 and constitutes a prominent antigenic domain that interacts directly with neutralizing antibodies. Previous studies have shown that this domain is also important for penetration of cells and for virulence of the virus. Resistance to the single E1-specific neutralizing monoclonal antibody resulted from changes of Gly-132 of E1 to either Arg or Glu. Analogous to the findings with E2, these changes result in a change in charge and are found near a glycosylation site at residue 139. This domain of E1 may therefore be found near the 181 to 216 domain of E2 on the surface of the E1-E2 heterodimer; together, they could form a domain important in virus penetration and neutralization.

Full text

PDF
4655

Images in this article

4662Image
on p.4662

Selected References

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

  1. Boere W. A., Benaissa-Trouw B. J., Harmsen T., Erich T., Kraaijeveld C. A., Snippe H. Mechanisms of monoclonal antibody-mediated protection against virulent Semliki Forest virus. J Virol. 1985 May;54(2):546–551. doi: 10.1128/jvi.54.2.546-551.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Burge B. W., Pfefferkorn E. R. Isolation and characterization of conditional-lethal mutants of Sindbis virus. Virology. 1966 Oct;30(2):204–213. doi: 10.1016/0042-6822(66)90096-1. [DOI] [PubMed] [Google Scholar]
  3. Chanas A. C., Johnson B. K., Simpson D. I. Antigenic relationships of alphaviruses by a simple micro-culture cross-neutralization method. J Gen Virol. 1976 Aug;32(2):295–300. doi: 10.1099/0022-1317-32-2-295. [DOI] [PubMed] [Google Scholar]
  4. Co M. S., Gaulton G. N., Fields B. N., Greene M. I. Isolation and biochemical characterization of the mammalian reovirus type 3 cell-surface receptor. Proc Natl Acad Sci U S A. 1985 Mar;82(5):1494–1498. doi: 10.1073/pnas.82.5.1494. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. DULBECCO R., VOGT M. One-step growth curve of Western equine encephalomyelitis virus on chicken embryo cells grown in vitro and analysis of virus yields from single cells. J Exp Med. 1954 Feb;99(2):183–199. doi: 10.1084/jem.99.2.183. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Davis N. L., Pence D. F., Meyer W. J., Schmaljohn A. L., Johnston R. E. Alternative forms of a strain-specific neutralizing antigenic site on the Sindbis virus E2 glycoprotein. Virology. 1987 Nov;161(1):101–108. doi: 10.1016/0042-6822(87)90175-9. [DOI] [PubMed] [Google Scholar]
  7. Durbin R. K., Stollar V. Sequence analysis of the E2 gene of a hyperglycosylated, host restricted mutant of Sindbis virus and estimation of mutation rate from frequency of revertants. Virology. 1986 Oct 15;154(1):135–143. doi: 10.1016/0042-6822(86)90436-8. [DOI] [PubMed] [Google Scholar]
  8. 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]
  9. Gaulton G. N., Greene M. I. Idiotypic mimicry of biological receptors. Annu Rev Immunol. 1986;4:253–280. doi: 10.1146/annurev.iy.04.040186.001345. [DOI] [PubMed] [Google Scholar]
  10. Hahn C. S., Strauss E. G., Strauss J. H. Dideoxy sequencing of RNA using reverse transcriptase. Methods Enzymol. 1989;180:121–130. doi: 10.1016/0076-6879(89)80097-7. [DOI] [PubMed] [Google Scholar]
  11. Johnson B. J., Brubaker J. R., Roehrig J. T., Trent D. W. Variants of Venezuelan equine encephalitis virus that resist neutralization define a domain of the E2 glycoprotein. Virology. 1990 Aug;177(2):676–683. doi: 10.1016/0042-6822(90)90533-w. [DOI] [PubMed] [Google Scholar]
  12. Keegan K., Collett M. S. Use of bacterial expression cloning to define the amino acid sequences of antigenic determinants on the G2 glycoprotein of Rift Valley fever virus. J Virol. 1986 May;58(2):263–270. doi: 10.1128/jvi.58.2.263-270.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
  14. Lindqvist B. H., DiSalvo J., Rice C. M., Strauss J. H., Strauss E. G. Sindbis virus mutant ts20 of complementation group E contains a lesion in glycoprotein E2. Virology. 1986 May;151(1):10–20. doi: 10.1016/0042-6822(86)90099-1. [DOI] [PubMed] [Google Scholar]
  15. Lobigs M., Dalgarno L., Schlesinger J. J., Weir R. C. Location of a neutralization determinant in the E protein of yellow fever virus (17D vaccine strain). Virology. 1987 Dec;161(2):474–478. doi: 10.1016/0042-6822(87)90141-3. [DOI] [PubMed] [Google Scholar]
  16. Lustig S., Jackson A. C., Hahn C. S., Griffin D. E., Strauss E. G., Strauss J. H. Molecular basis of Sindbis virus neurovirulence in mice. J Virol. 1988 Jul;62(7):2329–2336. doi: 10.1128/jvi.62.7.2329-2336.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Mathews J. H., Roehrig J. T. Determination of the protective epitopes on the glycoproteins of Venezuelan equine encephalomyelitis virus by passive transfer of monoclonal antibodies. J Immunol. 1982 Dec;129(6):2763–2767. [PubMed] [Google Scholar]
  18. Mäkelä M. J., Lund G. A., Salmi A. A. Antigenicity of the measles virus haemagglutinin studied by using synthetic peptides. J Gen Virol. 1989 Mar;70(Pt 3):603–614. doi: 10.1099/0022-1317-70-3-603. [DOI] [PubMed] [Google Scholar]
  19. 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]
  20. Ou J. H., Strauss E. G., Strauss J. H. Comparative studies of the 3'-terminal sequences of several alpha virus RNAs. Virology. 1981 Mar;109(2):281–289. doi: 10.1016/0042-6822(81)90499-2. [DOI] [PubMed] [Google Scholar]
  21. Parry N., Fox G., Rowlands D., Brown F., Fry E., Acharya R., Logan D., Stuart D. Structural and serological evidence for a novel mechanism of antigenic variation in foot-and-mouth disease virus. Nature. 1990 Oct 11;347(6293):569–572. doi: 10.1038/347569a0. [DOI] [PubMed] [Google Scholar]
  22. Pence D. F., Davis N. L., Johnston R. E. Antigenic and genetic characterization of Sindbis virus monoclonal antibody escape mutants which define a pathogenesis domain on glycoprotein E2. Virology. 1990 Mar;175(1):41–49. doi: 10.1016/0042-6822(90)90184-s. [DOI] [PubMed] [Google Scholar]
  23. Pierce J. S., Strauss E. G., Strauss J. H. Effect of ionic strength on the binding of Sindbis virus to chick cells. J Virol. 1974 May;13(5):1030–1036. doi: 10.1128/jvi.13.5.1030-1036.1974. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. 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]
  25. Rice C. M., Strauss J. H. Nucleotide sequence of the 26S mRNA of Sindbis virus and deduced sequence of the encoded virus structural proteins. Proc Natl Acad Sci U S A. 1981 Apr;78(4):2062–2066. doi: 10.1073/pnas.78.4.2062. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Roehrig J. T., Day J. W., Kinney R. M. Antigenic analysis of the surface glycoproteins of a Venezuelan equine encephalomyelitis virus (TC-83) using monoclonal antibodies. Virology. 1982 Apr 30;118(2):269–278. doi: 10.1016/0042-6822(82)90346-4. [DOI] [PubMed] [Google Scholar]
  27. 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]
  28. Roehrig J. T., Hunt A. R., Kinney R. M., Mathews J. H. In vitro mechanisms of monoclonal antibody neutralization of alphaviruses. Virology. 1988 Jul;165(1):66–73. doi: 10.1016/0042-6822(88)90659-9. [DOI] [PubMed] [Google Scholar]
  29. Roehrig J. T., Mathews J. H. The neutralization site on the E2 glycoprotein of Venezuelan equine encephalomyelitis (TC-83) virus is composed of multiple conformationally stable epitopes. Virology. 1985 Apr 30;142(2):347–356. doi: 10.1016/0042-6822(85)90343-5. [DOI] [PubMed] [Google Scholar]
  30. 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]
  31. Sanger F., Nicklen S., Coulson A. R. DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci U S A. 1977 Dec;74(12):5463–5467. doi: 10.1073/pnas.74.12.5463. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. 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]
  33. 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]
  34. Schmaljohn C. S., Schmaljohn A. L., Dalrymple J. M. Hantaan virus M RNA: coding strategy, nucleotide sequence, and gene order. Virology. 1987 Mar;157(1):31–39. doi: 10.1016/0042-6822(87)90310-2. [DOI] [PubMed] [Google Scholar]
  35. Sherry B., Mosser A. G., Colonno R. J., Rueckert R. R. Use of monoclonal antibodies to identify four neutralization immunogens on a common cold picornavirus, human rhinovirus 14. J Virol. 1986 Jan;57(1):246–257. doi: 10.1128/jvi.57.1.246-257.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Shirako Y., Niklasson B., Dalrymple J. M., Strauss E. G., Strauss J. H. Structure of the Ockelbo virus genome and its relationship to other Sindbis viruses. Virology. 1991 Jun;182(2):753–764. doi: 10.1016/0042-6822(91)90616-j. [DOI] [PubMed] [Google Scholar]
  37. Smith D. R., Calvo J. M. Nucleotide sequence of the E coli gene coding for dihydrofolate reductase. Nucleic Acids Res. 1980 May 24;8(10):2255–2274. doi: 10.1093/nar/8.10.2255. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. 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]
  39. Stanley J., Cooper S. J., Griffin D. E. Monoclonal antibody cure and prophylaxis of lethal Sindbis virus encephalitis in mice. J Virol. 1986 Apr;58(1):107–115. doi: 10.1128/jvi.58.1.107-115.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  40. Stec D. S., Waddell A., Schmaljohn C. S., Cole G. A., Schmaljohn A. L. Antibody-selected variation and reversion in Sindbis virus neutralization epitopes. J Virol. 1986 Mar;57(3):715–720. doi: 10.1128/jvi.57.3.715-720.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  41. 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]
  42. TAYLOR R. M., HURLBUT H. S., WORK T. H., KINGSTON J. R., FROTHINGHAM T. E. Sindbis virus: a newly recognized arthropodtransmitted virus. Am J Trop Med Hyg. 1955 Sep;4(5):844–862. doi: 10.4269/ajtmh.1955.4.844. [DOI] [PubMed] [Google Scholar]
  43. Tucker P. C., Griffin D. E. Mechanism of altered Sindbis virus neurovirulence associated with a single-amino-acid change in the E2 Glycoprotein. J Virol. 1991 Mar;65(3):1551–1557. doi: 10.1128/jvi.65.3.1551-1557.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  44. Vrati S., Fernon C. A., Dalgarno L., Weir R. C. Location of a major antigenic site involved in Ross River virus neutralization. Virology. 1988 Feb;162(2):346–353. doi: 10.1016/0042-6822(88)90474-6. [DOI] [PubMed] [Google Scholar]
  45. Wang K. S., Schmaljohn A. L., Kuhn R. J., Strauss J. H. Antiidiotypic antibodies as probes for the Sindbis virus receptor. Virology. 1991 Apr;181(2):694–702. doi: 10.1016/0042-6822(91)90903-o. [DOI] [PubMed] [Google Scholar]
  46. White J. M., Littman D. R. Viral receptors of the immunoglobulin superfamily. Cell. 1989 Mar 10;56(5):725–728. doi: 10.1016/0092-8674(89)90674-0. [DOI] [PubMed] [Google Scholar]
  47. Wiley D. C., Skehel J. J. The structure and function of the hemagglutinin membrane glycoprotein of influenza virus. Annu Rev Biochem. 1987;56:365–394. doi: 10.1146/annurev.bi.56.070187.002053. [DOI] [PubMed] [Google Scholar]
  48. Yamamoto K., Hashimoto K., Chiba J., Simizu B. Properties of monoclonal antibodies against glycoproteins of western equine encephalitis virus. J Virol. 1985 Sep;55(3):840–842. doi: 10.1128/jvi.55.3.840-842.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  49. Ziemiecki A., Garofff H. Subunit composition of the membrane glycoprotein complex of Semliki Forest virus. J Mol Biol. 1978 Jul 5;122(3):259–269. doi: 10.1016/0022-2836(78)90189-4. [DOI] [PubMed] [Google Scholar]
  50. Zimmern D., Kaesberg P. 3'-terminal nucleotide sequence of encephalomyocarditis virus RNA determined by reverse transcriptase and chain-terminating inhibitors. Proc Natl Acad Sci U S A. 1978 Sep;75(9):4257–4261. doi: 10.1073/pnas.75.9.4257. [DOI] [PMC free article] [PubMed] [Google Scholar]

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

RESOURCES