Skip to main content
Journal of Virology logoLink to Journal of Virology
. 1990 Mar;64(3):1044–1049. doi: 10.1128/jvi.64.3.1044-1049.1990

Rotavirus genome segment 4 determines viral replication phenotype in cultured liver cells (HepG2).

R F Ramig 1, K L Galle 1
PMCID: PMC249215  PMID: 2154590

Abstract

One-step growth determinations were performed with five strains of rotavirus in HepG2, a cell line derived from human liver. Three virus strains (SA11-C13, SA11-C14, and RRV) replicated in HepG2 cells and attained yields 10- to 100-fold above input titers. Two virus strains (B223 and SA11-4F) failed to replicate above input titer. Analysis of reassortants that segregated the genes of parental virus pairs able and unable to replicate revealed that the HepG2 cell growth phenotype segregated with genome segment 4. Immunofluorescence analysis of infected HepG2 cells showed that the production of detectable antigen correlated with the growth phenotype and also segregated with genome segment 4. Thus, we conclude that (i) some virus strains were capable of replication in cultured liver cells while other strains could not replicate under identical conditions and that (ii) the inability of some virus strains to replicate resulted from a segment 4-associated block in replication before protein synthesis. These results are discussed in terms of what is known of the functions of VP4.

Full text

PDF

Images in this article

Selected References

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

  1. Adams W. R., Kraft L. M. Electron-Microscopic Study of the Intestinal Epithelium of Mice Infected with the Agent of Epizootic Diarrhea of Infant Mice (EDIM Virus). Am J Pathol. 1967 Jul;51(1):39–60. [PMC free article] [PubMed] [Google Scholar]
  2. Aden D. P., Fogel A., Plotkin S., Damjanov I., Knowles B. B. Controlled synthesis of HBsAg in a differentiated human liver carcinoma-derived cell line. Nature. 1979 Dec 6;282(5739):615–616. doi: 10.1038/282615a0. [DOI] [PubMed] [Google Scholar]
  3. Bishop R. F., Davidson G. P., Holmes I. H., Ruck B. J. Virus particles in epithelial cells of duodenal mucosa from children with acute non-bacterial gastroenteritis. Lancet. 1973 Dec 8;2(7841):1281–1283. doi: 10.1016/s0140-6736(73)92867-5. [DOI] [PubMed] [Google Scholar]
  4. Bridger J. C., Woode G. N. Characterization of two particle types of calf rotavirus. J Gen Virol. 1976 May;31(2):245–250. doi: 10.1099/0022-1317-31-2-245. [DOI] [PubMed] [Google Scholar]
  5. Burns J. W., Chen D., Estes M. K., Ramig R. F. Biological and immunological characterization of a simian rotavirus SA11 variant with an altered genome segment 4. Virology. 1989 Apr;169(2):427–435. doi: 10.1016/0042-6822(89)90168-2. [DOI] [PubMed] [Google Scholar]
  6. Chen D., Burns J. W., Estes M. K., Ramig R. F. Phenotypes of rotavirus reassortants depend upon the recipient genetic background. Proc Natl Acad Sci U S A. 1989 May;86(10):3743–3747. doi: 10.1073/pnas.86.10.3743. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Darlington G. J., Kelly J. H., Buffone G. J. Growth and hepatospecific gene expression of human hepatoma cells in a defined medium. In Vitro Cell Dev Biol. 1987 May;23(5):349–354. doi: 10.1007/BF02620991. [DOI] [PubMed] [Google Scholar]
  8. Davidson G. P., Goller I., Bishop R. F., Townley R. R., Holmes I. H., Ruck B. J. Immunofluorescence in duodenal mucosa of children with acute enteritis due to a new virus. J Clin Pathol. 1975 Apr;28(4):263–266. doi: 10.1136/jcp.28.4.263. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Estes M. K., Graham D. Y., Mason B. B. Proteolytic enhancement of rotavirus infectivity: molecular mechanisms. J Virol. 1981 Sep;39(3):879–888. doi: 10.1128/jvi.39.3.879-888.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Estes M. K., Palmer E. L., Obijeski J. F. Rotaviruses: a review. Curr Top Microbiol Immunol. 1983;105:123–184. doi: 10.1007/978-3-642-69159-1_3. [DOI] [PubMed] [Google Scholar]
  11. Gombold J. L., Ramig R. F. Analysis of reassortment of genome segments in mice mixedly infected with rotaviruses SA11 and RRV. J Virol. 1986 Jan;57(1):110–116. doi: 10.1128/jvi.57.1.110-116.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Greenberg H. B., Kalica A. R., Wyatt R. G., Jones R. W., Kapikian A. Z., Chanock R. M. Rescue of noncultivatable human rotavirus by gene reassortment during mixed infection with ts mutants of a cultivatable bovine rotavirus. Proc Natl Acad Sci U S A. 1981 Jan;78(1):420–424. doi: 10.1073/pnas.78.1.420. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Grimwood K., Coakley J. C., Hudson I. L., Bishop R. F., Barnes G. L. Serum aspartate aminotransferase levels after rotavirus gastroenteritis. J Pediatr. 1988 Apr;112(4):597–600. doi: 10.1016/s0022-3476(88)80179-3. [DOI] [PubMed] [Google Scholar]
  14. Grunow J. E., Dunton S. F., Waner J. L. Human rotavirus-like particles in a hepatic abscess. J Pediatr. 1985 Jan;106(1):73–76. doi: 10.1016/s0022-3476(85)80470-4. [DOI] [PubMed] [Google Scholar]
  15. Hoshino Y., Sereno M. M., Midthun K., Flores J., Kapikian A. Z., Chanock R. M. Independent segregation of two antigenic specificities (VP3 and VP7) involved in neutralization of rotavirus infectivity. Proc Natl Acad Sci U S A. 1985 Dec;82(24):8701–8704. doi: 10.1073/pnas.82.24.8701. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Johnson M. A., McCrae M. A. Molecular biology of rotaviruses. VIII. Quantitative analysis of regulation of gene expression during virus replication. J Virol. 1989 May;63(5):2048–2055. doi: 10.1128/jvi.63.5.2048-2055.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. KRAFT L. M. Observations on the control and natural history of epidemic diarrhea of infant mice (EDIM). Yale J Biol Med. 1958 Dec;31(3):121–137. [PMC free article] [PubMed] [Google Scholar]
  18. Kalica A. R., Flores J., Greenberg H. B. Identification of the rotaviral gene that codes for hemagglutination and protease-enhanced plaque formation. Virology. 1983 Feb;125(1):194–205. doi: 10.1016/0042-6822(83)90073-9. [DOI] [PubMed] [Google Scholar]
  19. Kaljot K. T., Shaw R. D., Rubin D. H., Greenberg H. B. Infectious rotavirus enters cells by direct cell membrane penetration, not by endocytosis. J Virol. 1988 Apr;62(4):1136–1144. doi: 10.1128/jvi.62.4.1136-1144.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Kovacs A., Chan L., Hotrakitya C., Overturf G., Portnoy B. Serum transaminase elevations in infants with rotavirus gastroenteritis. J Pediatr Gastroenterol Nutr. 1986 Nov-Dec;5(6):873–877. doi: 10.1097/00005176-198611000-00008. [DOI] [PubMed] [Google Scholar]
  21. McAdaragh J. P., Bergeland M. E., Meyer R. C., Johnshoy M. W., Stotz I. J., Benfield D. A., Hammer R. Pathogenesis of rotaviral enteritis in gnotobiotic pigs: a microscopic study. Am J Vet Res. 1980 Oct;41(10):1572–1581. [PubMed] [Google Scholar]
  22. Offit P. A., Blavat G. Identification of the two rotavirus genes determining neutralization specificities. J Virol. 1986 Jan;57(1):376–378. doi: 10.1128/jvi.57.1.376-378.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Pearson G. R., McNulty M. S., Logan E. F. Pathological changes in the small intestine of neonatal calves naturally infected with reo-like virus (rotavirus). Vet Rec. 1978 May 27;102(21):454–458. doi: 10.1136/vr.102.21.454. [DOI] [PubMed] [Google Scholar]
  24. Prasad B. V., Wang G. J., Clerx J. P., Chiu W. Three-dimensional structure of rotavirus. J Mol Biol. 1988 Jan 20;199(2):269–275. doi: 10.1016/0022-2836(88)90313-0. [DOI] [PubMed] [Google Scholar]
  25. Ramig R. F. Isolation and genetic characterization of temperature-sensitive mutants of simian rotavirus SA11. Virology. 1982 Jul 15;120(1):93–105. doi: 10.1016/0042-6822(82)90009-5. [DOI] [PubMed] [Google Scholar]
  26. Ramig R. F. Isolation and genetic characterization of temperature-sensitive mutants of simian rotavirus SA11. Virology. 1982 Jul 15;120(1):93–105. doi: 10.1016/0042-6822(82)90009-5. [DOI] [PubMed] [Google Scholar]
  27. Riepenhoff-Talty M., Dharakul T., Kowalski E., Michalak S., Ogra P. L. Persistent rotavirus infection in mice with severe combined immunodeficiency. J Virol. 1987 Oct;61(10):3345–3348. doi: 10.1128/jvi.61.10.3345-3348.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Sabara M., Gilchrist J. E., Hudson G. R., Babiuk L. A. Preliminary characterization of an epitope involved in neutralization and cell attachment that is located on the major bovine rotavirus glycoprotein. J Virol. 1985 Jan;53(1):58–66. doi: 10.1128/jvi.53.1.58-66.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Sharpe A. H., Fields B. N. Reovirus inhibition of cellular RNA and protein synthesis: role of the S4 gene. Virology. 1982 Oct 30;122(2):381–391. doi: 10.1016/0042-6822(82)90237-9. [DOI] [PubMed] [Google Scholar]
  30. Tallett S., MacKenzie C., Middleton P., Kerzner B., Hamilton R. Clinical, laboratory, and epidemiologic features of a viral gastroenteritis in infants and children. Pediatrics. 1977 Aug;60(2):217–222. [PubMed] [Google Scholar]

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

RESOURCES