Abstract
Astroviruses are nonenveloped particles with a distinctive star-shaped surface structure that have been detected by electron microscopy in stool samples from humans and animals with gastroenteritis. We examined the patterns of macromolecular synthesis in astrovirus-infected cells with a goal of establishing a molecular basis for taxonomic classification. Trypsin is required for continuous replication of astrovirus in cultured cells; however, during a single cycle of infection, astrovirus antigen was synthesized earlier and at higher levels when serum, rather than trypsin, was included in the growth medium. This enhanced production of antigen, as measured by enzyme immunoassay, was accompanied by the appearance of aggregates of virus particles in the cytoplasm of infected cells. During astrovirus replication in cells cultured in the presence of serum, we detected a single infection-specific protein (90 kDa) beginning at 12 h postinfection. This protein was recognized by antiastrovirus rabbit serum and was sensitive to trypsin digestion in vitro, with the concomitant appearance of three smaller immunoreactive proteins (31, 29, and 20 kDa). We also detected two dactinomycin-resistant RNAs (7.2 and 2.8 kb), both of which were polyadenylated, in the cytoplasm of astrovirus-infected cells. The larger of these two RNAs is presumably the viral genome, whereas the smaller species may be a subgenomic messenger. Comparison of the proteins and RNAs synthesized in astrovirus-infected cells with those of the recognized families of nonenveloped single-stranded RNA animal viruses suggests that astroviruses should not be classified as members of either Caliciviridae or Picornaviridae.
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- Aroonprasert D., Fagerland J. A., Kelso N. E., Zheng S., Woode G. N. Cultivation and partial characterization of bovine astrovirus. Vet Microbiol. 1989 Feb;19(2):113–125. doi: 10.1016/0378-1135(89)90077-1. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Ashley C. R., Caul E. O., Paver W. K. Astrovirus-associated gastroenteritis in children. J Clin Pathol. 1978 Oct;31(10):939–943. doi: 10.1136/jcp.31.10.939. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Carter M. J. Feline calicivirus protein synthesis investigated by western blotting. Arch Virol. 1989;108(1-2):69–79. doi: 10.1007/BF01313744. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Chomczynski P., Sacchi N. Single-step method of RNA isolation by acid guanidinium thiocyanate-phenol-chloroform extraction. Anal Biochem. 1987 Apr;162(1):156–159. doi: 10.1006/abio.1987.9999. [DOI] [PubMed] [Google Scholar]
- Cubitt W. D., Barrett A. D. Propagation of human candidate calicivirus in cell culture. J Gen Virol. 1984 Jun;65(Pt 6):1123–1126. doi: 10.1099/0022-1317-65-6-1123. [DOI] [PubMed] [Google Scholar]
- 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]
- Flynn W. T., Saif L. J. Serial propagation of porcine enteric calicivirus-like virus in primary porcine kidney cell cultures. J Clin Microbiol. 1988 Feb;26(2):206–212. doi: 10.1128/jcm.26.2.206-212.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Gray J. J., Wreghitt T. G., Cubitt W. D., Elliot P. R. An outbreak of gastroenteritis in a home for the elderly associated with astrovirus type 1 and human calicivirus. J Med Virol. 1987 Dec;23(4):377–381. doi: 10.1002/jmv.1890230410. [DOI] [PubMed] [Google Scholar]
- Harbour D. A., Ashley C. R., Williams P. D., Gruffydd-Jones T. J. Natural and experimental astrovirus infection of cats. Vet Rec. 1987 Jun 6;120(23):555–557. doi: 10.1136/vr.120.23.555. [DOI] [PubMed] [Google Scholar]
- Herring A. J., Gray E. W., Snodgrass D. R. Purification and characterization of ovine astrovirus. J Gen Virol. 1981 Mar;53(Pt 1):47–55. doi: 10.1099/0022-1317-53-1-47. [DOI] [PubMed] [Google Scholar]
- Herrmann J. E., Cubitt W. D., Hudson R. W., Perron-Henry D. M., Oshiro L. S., Blacklow N. R. Immunological characterization of the Marin County strain of astrovirus. Arch Virol. 1990;110(3-4):213–220. doi: 10.1007/BF01311289. [DOI] [PubMed] [Google Scholar]
- Herrmann J. E., Hudson R. W., Blacklow N. R. Marin County agent, an astrovirus. Lancet. 1987 Sep 26;2(8561):743–743. doi: 10.1016/s0140-6736(87)91102-0. [DOI] [PubMed] [Google Scholar]
- Herrmann J. E., Hudson R. W., Perron-Henry D. M., Kurtz J. B., Blacklow N. R. Antigenic characterization of cell-cultivated astrovirus serotypes and development of astrovirus-specific monoclonal antibodies. J Infect Dis. 1988 Jul;158(1):182–185. doi: 10.1093/infdis/158.1.182. [DOI] [PubMed] [Google Scholar]
- Herrmann J. E., Nowak N. A., Perron-Henry D. M., Hudson R. W., Cubitt W. D., Blacklow N. R. Diagnosis of astrovirus gastroenteritis by antigen detection with monoclonal antibodies. J Infect Dis. 1990 Feb;161(2):226–229. doi: 10.1093/infdis/161.2.226. [DOI] [PubMed] [Google Scholar]
- Hudson R. W., Herrmann J. E., Blacklow N. R. Plaque quantitation and virus neutralization assays for human astroviruses. Arch Virol. 1989;108(1-2):33–38. doi: 10.1007/BF01313740. [DOI] [PubMed] [Google Scholar]
- Konno T., Suzuki H., Ishida N., Chiba R., Mochizuki K., Tsunoda A. Astrovirus-associated epidemic gastroenteritis in Japan. J Med Virol. 1982;9(1):11–17. doi: 10.1002/jmv.1890090103. [DOI] [PubMed] [Google Scholar]
- Kurtz J. B., Lee T. W. Astroviruses: human and animal. Ciba Found Symp. 1987;128:92–107. doi: 10.1002/9780470513460.ch6. [DOI] [PubMed] [Google Scholar]
- Kurtz J. B., Lee T. W., Craig J. W., Reed S. E. Astrovirus infection in volunteers. J Med Virol. 1979;3(3):221–230. doi: 10.1002/jmv.1890030308. [DOI] [PubMed] [Google Scholar]
- Kurtz J., Lee T. Astrovirus gastroenteritis age distribution of antibody. Med Microbiol Immunol. 1978 Nov 17;166(1-4):227–230. doi: 10.1007/BF02121154. [DOI] [PubMed] [Google Scholar]
- Lee T. W., Kurtz J. B. Human astrovirus serotypes. J Hyg (Lond) 1982 Dec;89(3):539–540. doi: 10.1017/s0022172400071114. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Lee T. W., Kurtz J. B. Serial propagation of astrovirus in tissue culture with the aid of trypsin. J Gen Virol. 1981 Dec;57(Pt 2):421–424. doi: 10.1099/0022-1317-57-2-421. [DOI] [PubMed] [Google Scholar]
- Madeley C. R. Comparison of the features of astroviruses and caliciviruses seen in samples of feces by electron microscopy. J Infect Dis. 1979 May;139(5):519–523. doi: 10.1093/infdis/139.5.519. [DOI] [PubMed] [Google Scholar]
- Madeley C. R., Cosgrove B. P. Letter: 28 nm particles in faeces in infantile gastroenteritis. Lancet. 1975 Sep 6;2(7932):451–452. doi: 10.1016/s0140-6736(75)90858-2. [DOI] [PubMed] [Google Scholar]
- Neill J. D., Mengeling W. L. Further characterization of the virus-specific RNAs in feline calicivirus infected cells. Virus Res. 1988 Aug;11(1):59–72. doi: 10.1016/0168-1702(88)90067-6. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Sanchez A., Kiley M. P. Identification and analysis of Ebola virus messenger RNA. Virology. 1987 Apr;157(2):414–420. doi: 10.1016/0042-6822(87)90283-2. [DOI] [PubMed] [Google Scholar]
- Shimizu M., Shirai J., Narita M., Yamane T. Cytopathic astrovirus isolated from porcine acute gastroenteritis in an established cell line derived from porcine embryonic kidney. J Clin Microbiol. 1990 Feb;28(2):201–206. doi: 10.1128/jcm.28.2.201-206.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Terrett L. A., Saif L. J. Serial propagation of porcine group C rotavirus (pararotavirus) in primary porcine kidney cell cultures. J Clin Microbiol. 1987 Jul;25(7):1316–1319. doi: 10.1128/jcm.25.7.1316-1319.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Urasawa T., Urasawa S., Taniguchi K. Sequential passages of human rotavirus in MA-104 cells. Microbiol Immunol. 1981;25(10):1025–1035. doi: 10.1111/j.1348-0421.1981.tb00109.x. [DOI] [PubMed] [Google Scholar]
- Wilson S. A., Cubitt W. D. The development and evaluation of radioimmune assays for the detection of immune globulins M and G against astrovirus. J Virol Methods. 1988 Feb;19(2):151–159. doi: 10.1016/0166-0934(88)90158-9. [DOI] [PubMed] [Google Scholar]
- Woode G. N., Bridger J. C. Isolation of small viruses resembling astroviruses and caliciviruses from acute enteritis of calves. J Med Microbiol. 1978 Nov;11(4):441–452. doi: 10.1099/00222615-11-4-441. [DOI] [PubMed] [Google Scholar]