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. 1989 May;63(5):2191–2197. doi: 10.1128/jvi.63.5.2191-2197.1989

Purification and characterization of adult diarrhea rotavirus: identification of viral structural proteins.

Z Y Fang 1, R I Glass 1, M Penaranda 1, H Dong 1, S S Monroe 1, L Wen 1, M K Estes 1, J Eiden 1, R H Yolken 1, L Saif 1, et al.
PMCID: PMC250636  PMID: 2539512

Abstract

Adult diarrhea rotavirus (ADRV) is a newly identified strain of noncultivable human group B rotavirus that has been epidemic in the People's Republic of China since 1982. We have used sodium dodecyl sulfate-polyacrylamide gel electrophoresis and Western (immuno-) blot analysis to examine the viral proteins present in the outer and inner capsids of ADRV and compared these with the proteins of a group A rotavirus, SA11. EDTA treatment of double-shelled virions removed the outer capsid and resulted in the loss of three polypeptides of 64, 61, and 41, kilodaltons (kDa). Endo-beta-N-acetylglucosaminidase H digestion of double-shelled virions identified the 41-kDa polypeptide as a glycoprotein. CaCl2 treatment of single-shelled particles removed the inner capsid and resulted in the loss of one polypeptide with a molecular mass of 47 kDa. The remaining core particle had two major structural proteins of 136 and 113 kDa. All of the proteins visualized on sodium dodecyl sulfate-polyacrylamide gel electrophoresis were antigenic by Western blot analysis when probed with convalescent-phase human and animal antisera. A 47-kDa polypeptide was most abundant and was strongly immunoreactive with human sera, animal sera raised against ADRV and against other group B animal rotaviruses (infectious diarrhea of infant rat virus, bovine and porcine group B rotavirus, and bovine enteric syncytial virus) and a monoclonal antibody prepared against infectious diarrhea of infant rat virus. This 47-kDa inner capsid polypeptide contains a common group B antigen and is similar to the VP6 of the group A rotaviruses. Human convalescent-phase sera also responded to a 41-kDa polypeptide of the outer capsid that seems similar to the VP7 of group A rotavirus. Other polypeptides have been given tentative designations on the basis of similarities to the control preparation of SA11, including a 136-kDa polypeptide designated VP1, a 113-kDa polypeptide designated VP2, 64- and 61-kDa polypeptides designated VP5 and VP5a, and several proteins in the 110- to 72-kDa range that may be VP3, VP4, or related proteins. The lack of cross-reactivity on Western blots between antisera to group A versus group B rotaviruses confirmed that these viruses are antigenically quite distinct.

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Selected References

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

  1. Bican P., Cohen J., Charpilienne A., Scherrer R. Purification and characterization of bovine rotavirus cores. J Virol. 1982 Sep;43(3):1113–1117. doi: 10.1128/jvi.43.3.1113-1117.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Bremont M., Cohen J., McCrae M. A. Analysis of the structural polypeptides of a porcine group C rotavirus. J Virol. 1988 Jun;62(6):2183–2185. doi: 10.1128/jvi.62.6.2183-2185.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Bridger J. C. Novel rotaviruses in animals and man. Ciba Found Symp. 1987;128:5–23. doi: 10.1002/9780470513460.ch2. [DOI] [PubMed] [Google Scholar]
  4. Bridger J. C., Pedley S., McCrae M. A. Group C rotaviruses in humans. J Clin Microbiol. 1986 Apr;23(4):760–763. doi: 10.1128/jcm.23.4.760-763.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Burns J. W., Welch S. K., Nakata S., Estes M. K. Characterization of monoclonal antibodies to human group B rotavirus and their use in an antigen detection enzyme-linked immunosorbent assay. J Clin Microbiol. 1989 Feb;27(2):245–250. doi: 10.1128/jcm.27.2.245-250.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Dimitrov D. H., Estes M. K., Rangelova S. M., Shindarov L. M., Melnick J. L., Graham D. Y. Detection of antigenically distinct rotaviruses from infants. Infect Immun. 1983 Aug;41(2):523–526. doi: 10.1128/iai.41.2.523-526.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Eiden J., Vonderfecht S., Theil K., Torres-Medina A., Yolken R. H. Genetic and antigenic relatedness of human and animal strains of antigenically distinct rotaviruses. J Infect Dis. 1986 Dec;154(6):972–982. doi: 10.1093/infdis/154.6.972. [DOI] [PubMed] [Google Scholar]
  8. Espejo R. T., Puerto F., Soler C., González N. Characterization of a human pararotavirus. Infect Immun. 1984 Apr;44(1):112–116. doi: 10.1128/iai.44.1.112-116.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Estes M. K., Graham D. Y., Gerba C. P., Smith E. M. Simian rotavirus SA11 replication in cell cultures. J Virol. 1979 Sep;31(3):810–815. doi: 10.1128/jvi.31.3.810-815.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Fang Z. Y., Deng S. S., Wang C. A., Ye W. W., Chen G. M., Chi J., Hong T. [Analysis of the genome of rotavirus from the feces of adult infectious diarrhea]. Zhongguo Yi Xue Ke Xue Yuan Xue Bao. 1985 Apr;7(2):93–96. [PubMed] [Google Scholar]
  11. Hung T., Chen G. M., Wang C. G., Yao H. L., Fang Z. Y., Chao T. X., Chou Z. Y., Ye W., Chang X. J., Den S. S. Waterborne outbreak of rotavirus diarrhoea in adults in China caused by a novel rotavirus. Lancet. 1984 May 26;1(8387):1139–1142. [PubMed] [Google Scholar]
  12. 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]
  13. Mason B. B., Graham D. Y., Estes M. K. Biochemical mapping of the simian rotavirus SA11 genome. J Virol. 1983 May;46(2):413–423. doi: 10.1128/jvi.46.2.413-423.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Mason B. B., Graham D. Y., Estes M. K. In vitro transcription and translation of simian rotavirus SA11 gene products. J Virol. 1980 Mar;33(3):1111–1121. doi: 10.1128/jvi.33.3.1111-1121.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Nakata S., Estes M. K., Graham D. Y., Loosle R., Tao H., Wang S. H., Saif L. J., Melnick J. L. Antigenic characterization and ELISA detection of adult diarrhea rotaviruses. J Infect Dis. 1986 Sep;154(3):448–455. doi: 10.1093/infdis/154.3.448. [DOI] [PubMed] [Google Scholar]
  16. Nakata S., Petrie B. L., Calomeni E. P., Estes M. K. Electron microscopy procedure influences detection of rotaviruses. J Clin Microbiol. 1987 Oct;25(10):1902–1906. doi: 10.1128/jcm.25.10.1902-1906.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Nicolas J. C., Cohen J., Fortier B., Lourenco M. H., Bricout F. Isolation of a human pararotavirus. Virology. 1983 Jan 15;124(1):181–184. doi: 10.1016/0042-6822(83)90302-1. [DOI] [PubMed] [Google Scholar]
  18. Pedley S., Bridger J. C., Brown J. F., McCrae M. A. Molecular characterization of rotaviruses with distinct group antigens. J Gen Virol. 1983 Oct;64(Pt 10):2093–2101. doi: 10.1099/0022-1317-64-10-2093. [DOI] [PubMed] [Google Scholar]
  19. Tao H. Rotavirus and adult diarrhea. Adv Virus Res. 1988;35:193–218. [PubMed] [Google Scholar]
  20. Theil K. W., Saif L. J., Moorhead P. D., Whitmoyer R. E. Porcine rotavirus-like virus (group B rotavirus): characterization and pathogenicity for gnotobiotic pigs. J Clin Microbiol. 1985 Mar;21(3):340–345. doi: 10.1128/jcm.21.3.340-345.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Towbin H., Staehelin T., Gordon J. Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proc Natl Acad Sci U S A. 1979 Sep;76(9):4350–4354. doi: 10.1073/pnas.76.9.4350. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Vonderfecht S. L., Eiden J. J., Torres A., Miskuff R. L., Mebus C. A., Yolken R. H. Identification of a bovine enteric syncytial virus as a nongroup A rotavirus. Am J Vet Res. 1986 Sep;47(9):1913–1918. [PubMed] [Google Scholar]
  23. Yolken R., Wee S. B., Eiden J., Kinney J., Vonderfecht S. Identification of a group-reactive epitope of group B rotaviruses recognized by monoclonal antibody and application to the development of a sensitive immunoassay for viral characterization. J Clin Microbiol. 1988 Sep;26(9):1853–1858. doi: 10.1128/jcm.26.9.1853-1858.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]

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