Skip to main content
PMC home page
Journal of Virology logoLink to Journal of Virology
. 1997 Mar;71(3):1834–1841. doi: 10.1128/jvi.71.3.1834-1841.1997

Mutations in type 3 reovirus that determine binding to sialic acid are contained in the fibrous tail domain of viral attachment protein sigma1.

J D Chappell 1, V L Gunn 1, J D Wetzel 1, G S Baer 1, T S Dermody 1
PMCID: PMC191253  PMID: 9032313

Abstract

The reovirus attachment protein, sigma1, determines numerous aspects of reovirus-induced disease, including viral virulence, pathways of spread, and tropism for certain types of cells in the central nervous system. The sigma1 protein projects from the virion surface and consists of two distinct morphologic domains, a virion-distal globular domain known as the head and an elongated fibrous domain, termed the tail, which is anchored into the virion capsid. To better understand structure-function relationships of sigma1 protein, we conducted experiments to identify sequences in sigma1 important for viral binding to sialic acid, a component of the receptor for type 3 reovirus. Three serotype 3 reovirus strains incapable of binding sialylated receptors were adapted to growth in murine erythroleukemia (MEL) cells, in which sialic acid is essential for reovirus infectivity. MEL-adapted (MA) mutant viruses isolated by serial passage in MEL cells acquired the capacity to bind sialic acid-containing receptors and demonstrated a dependence on sialic acid for infection of MEL cells. Analysis of reassortant viruses isolated from crosses of an MA mutant virus and a reovirus strain that does not bind sialic acid indicated that the sigma1 protein is solely responsible for efficient growth of MA mutant viruses in MEL cells. The deduced sigma1 amino acid sequences of the MA mutant viruses revealed that each strain contains a substitution within a short region of sequence in the sigma1 tail predicted to form beta-sheet. These studies identify specific sequences that determine the capacity of reovirus to bind sialylated receptors and suggest a location for a sialic acid-binding domain. Furthermore, the results support a model in which type 3 sigma1 protein contains discrete receptor binding domains, one in the head and another in the tail that binds sialic acid.

Full Text

The Full Text of this article is available as a PDF (129.3 KB).

Selected References

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

  1. Armstrong G. D., Paul R. W., Lee P. W. Studies on reovirus receptors of L cells: virus binding characteristics and comparison with reovirus receptors of erythrocytes. Virology. 1984 Oct 15;138(1):37–48. doi: 10.1016/0042-6822(84)90145-4. [DOI] [PubMed] [Google Scholar]
  2. Barbis D. P., Chang S. F., Parrish C. R. Mutations adjacent to the dimple of the canine parvovirus capsid structure affect sialic acid binding. Virology. 1992 Nov;191(1):301–308. doi: 10.1016/0042-6822(92)90192-r. [DOI] [PubMed] [Google Scholar]
  3. Bassel-Duby R., Spriggs D. R., Tyler K. L., Fields B. N. Identification of attenuating mutations on the reovirus type 3 S1 double-stranded RNA segment with a rapid sequencing technique. J Virol. 1986 Oct;60(1):64–67. doi: 10.1128/jvi.60.1.64-67.1986. [DOI] [PMC free article] [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. Dermody T. S., Nibert M. L., Bassel-Duby R., Fields B. N. A sigma 1 region important for hemagglutination by serotype 3 reovirus strains. J Virol. 1990 Oct;64(10):5173–5176. doi: 10.1128/jvi.64.10.5173-5176.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Dichter M. A., Weiner H. L., Fields B. N., Mitchell G., Noseworthy J., Gaulton G., Greene M. Antiidiotypic antibody to reovirus binds to neurons and protects from viral infection. Ann Neurol. 1986 Jun;19(6):555–558. doi: 10.1002/ana.410190606. [DOI] [PubMed] [Google Scholar]
  7. Dichter M. A., Weiner H. L. Infection of neuronal cell cultures with reovirus mimics in vitro patterns of neurotropism. Ann Neurol. 1984 Nov;16(5):603–610. doi: 10.1002/ana.410160512. [DOI] [PubMed] [Google Scholar]
  8. Dryden K. A., Wang G., Yeager M., Nibert M. L., Coombs K. M., Furlong D. B., Fields B. N., Baker T. S. Early steps in reovirus infection are associated with dramatic changes in supramolecular structure and protein conformation: analysis of virions and subviral particles by cryoelectron microscopy and image reconstruction. J Cell Biol. 1993 Sep;122(5):1023–1041. doi: 10.1083/jcb.122.5.1023. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Duncan R., Horne D., Strong J. E., Leone G., Pon R. T., Yeung M. C., Lee P. W. Conformational and functional analysis of the C-terminal globular head of the reovirus cell attachment protein. Virology. 1991 Jun;182(2):810–819. doi: 10.1016/0042-6822(91)90622-i. [DOI] [PubMed] [Google Scholar]
  10. Fraser R. D., Furlong D. B., Trus B. L., Nibert M. L., Fields B. N., Steven A. C. Molecular structure of the cell-attachment protein of reovirus: correlation of computer-processed electron micrographs with sequence-based predictions. J Virol. 1990 Jun;64(6):2990–3000. doi: 10.1128/jvi.64.6.2990-3000.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. 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]
  12. Graessmann M., Graessmann A. Microinjection of tissue culture cells. Methods Enzymol. 1983;101:482–492. doi: 10.1016/0076-6879(83)01033-2. [DOI] [PubMed] [Google Scholar]
  13. Kowalik T. F., Yang Y. Y., Li J. K. Molecular cloning and comparative sequence analyses of bluetongue virus S1 segments by selective synthesis of specific full-length DNA copies of dsRNA genes. Virology. 1990 Aug;177(2):820–823. doi: 10.1016/0042-6822(90)90557-8. [DOI] [PubMed] [Google Scholar]
  14. Méndez E., Arias C. F., López S. Binding to sialic acids is not an essential step for the entry of animal rotaviruses to epithelial cells in culture. J Virol. 1993 Sep;67(9):5253–5259. doi: 10.1128/jvi.67.9.5253-5259.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Nagata L., Masri S. A., Pon R. T., Lee P. W. Analysis of functional domains on reovirus cell attachment protein sigma 1 using cloned S1 gene deletion mutants. Virology. 1987 Sep;160(1):162–168. doi: 10.1016/0042-6822(87)90056-0. [DOI] [PubMed] [Google Scholar]
  16. Nibert M. L., Chappell J. D., Dermody T. S. Infectious subvirion particles of reovirus type 3 Dearing exhibit a loss in infectivity and contain a cleaved sigma 1 protein. J Virol. 1995 Aug;69(8):5057–5067. doi: 10.1128/jvi.69.8.5057-5067.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Nibert M. L., Dermody T. S., Fields B. N. Structure of the reovirus cell-attachment protein: a model for the domain organization of sigma 1. J Virol. 1990 Jun;64(6):2976–2989. doi: 10.1128/jvi.64.6.2976-2989.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Pacitti A. F., Gentsch J. R. Inhibition of reovirus type 3 binding to host cells by sialylated glycoproteins is mediated through the viral attachment protein. J Virol. 1987 May;61(5):1407–1415. doi: 10.1128/jvi.61.5.1407-1415.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Paul R. W., Lee P. W. Glycophorin is the reovirus receptor on human erythrocytes. Virology. 1987 Jul;159(1):94–101. doi: 10.1016/0042-6822(87)90351-5. [DOI] [PubMed] [Google Scholar]
  20. ROSEN L., ABINANTI F. R., HOVIS J. F. Further observations on the natural infection of cattle with reoviruses. Am J Hyg. 1963 Jan;77:38–48. doi: 10.1093/oxfordjournals.aje.a120294. [DOI] [PubMed] [Google Scholar]
  21. ROSEN L. Serologic grouping of reoviruses by hemagglutination-inhibition. Am J Hyg. 1960 Mar;71:242–249. doi: 10.1093/oxfordjournals.aje.a120107. [DOI] [PubMed] [Google Scholar]
  22. Rubin D. H., Wetzel J. D., Williams W. V., Cohen J. A., Dworkin C., Dermody T. S. Binding of type 3 reovirus by a domain of the sigma 1 protein important for hemagglutination leads to infection of murine erythroleukemia cells. J Clin Invest. 1992 Dec;90(6):2536–2542. doi: 10.1172/JCI116147. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Saragovi H. U., Fitzpatrick D., Raktabutr A., Nakanishi H., Kahn M., Greene M. I. Design and synthesis of a mimetic from an antibody complementarity-determining region. Science. 1991 Aug 16;253(5021):792–795. doi: 10.1126/science.1876837. [DOI] [PubMed] [Google Scholar]
  24. Spriggs D. R., Bronson R. T., Fields B. N. Hemagglutinin variants of reovirus type 3 have altered central nervous system tropism. Science. 1983 Apr 29;220(4596):505–507. doi: 10.1126/science.6301010. [DOI] [PubMed] [Google Scholar]
  25. Tardieu M., Weiner H. L. Viral receptors on isolated murine and human ependymal cells. Science. 1982 Jan 22;215(4531):419–421. doi: 10.1126/science.6276976. [DOI] [PubMed] [Google Scholar]
  26. Tresnan D. B., Southard L., Weichert W., Sgro J. Y., Parrish C. R. Analysis of the cell and erythrocyte binding activities of the dimple and canyon regions of the canine parvovirus capsid. Virology. 1995 Aug 1;211(1):123–132. doi: 10.1006/viro.1995.1385. [DOI] [PubMed] [Google Scholar]
  27. Turner D. L., Duncan R., Lee P. W. Site-directed mutagenesis of the C-terminal portion of reovirus protein sigma 1: evidence for a conformation-dependent receptor binding domain. Virology. 1992 Jan;186(1):219–227. doi: 10.1016/0042-6822(92)90076-2. [DOI] [PubMed] [Google Scholar]
  28. Tyler K. L., McPhee D. A., Fields B. N. Distinct pathways of viral spread in the host determined by reovirus S1 gene segment. Science. 1986 Aug 15;233(4765):770–774. doi: 10.1126/science.3016895. [DOI] [PubMed] [Google Scholar]
  29. Virgin H. W., 4th, Bassel-Duby R., Fields B. N., Tyler K. L. Antibody protects against lethal infection with the neurally spreading reovirus type 3 (Dearing). J Virol. 1988 Dec;62(12):4594–4604. doi: 10.1128/jvi.62.12.4594-4604.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Weiner H. L., Drayna D., Averill D. R., Jr, Fields B. N. Molecular basis of reovirus virulence: role of the S1 gene. Proc Natl Acad Sci U S A. 1977 Dec;74(12):5744–5748. doi: 10.1073/pnas.74.12.5744. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Weiner H. L., Fields B. N. Neutralization of reovirus: the gene responsible for the neutralization antigen. J Exp Med. 1977 Nov 1;146(5):1305–1310. doi: 10.1084/jem.146.5.1305. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Weiner H. L., Powers M. L., Fields B. N. Absolute linkage of virulence and central nervous system cell tropism of reoviruses to viral hemagglutinin. J Infect Dis. 1980 May;141(5):609–616. doi: 10.1093/infdis/141.5.609. [DOI] [PubMed] [Google Scholar]
  33. Wickham T. J., Mathias P., Cheresh D. A., Nemerow G. R. Integrins alpha v beta 3 and alpha v beta 5 promote adenovirus internalization but not virus attachment. Cell. 1993 Apr 23;73(2):309–319. doi: 10.1016/0092-8674(93)90231-e. [DOI] [PubMed] [Google Scholar]
  34. Williams W. V., Guy H. R., Rubin D. H., Robey F., Myers J. N., Kieber-Emmons T., Weiner D. B., Greene M. I. Sequences of the cell-attachment sites of reovirus type 3 and its anti-idiotypic/antireceptor antibody: modeling of their three-dimensional structures. Proc Natl Acad Sci U S A. 1988 Sep;85(17):6488–6492. doi: 10.1073/pnas.85.17.6488. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Yeung M. C., Lim D., Duncan R., Shahrabadi M. S., Cashdollar L. W., Lee P. W. The cell attachment proteins of type 1 and type 3 reovirus are differentially susceptible to trypsin and chymotrypsin. Virology. 1989 May;170(1):62–70. doi: 10.1016/0042-6822(89)90352-8. [DOI] [PubMed] [Google Scholar]
  36. Zhou Y. J., Burns J. W., Morita Y., Tanaka T., Estes M. K. Localization of rotavirus VP4 neutralization epitopes involved in antibody-induced conformational changes of virus structure. J Virol. 1994 Jun;68(6):3955–3964. doi: 10.1128/jvi.68.6.3955-3964.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]

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

RESOURCES