Abstract
Successful viral infection involves a series of interactions between the virus and the host cell. The outcome of viral infection is, in fact, dependent on intact cellular function; it is required for viral binding, internalization, and uncoating. To determine the potential importance of lysosomal processing on the outcome of infection with a nonenveloped virus, we have studied the effects of NH4Cl on the course of reovirus infection on a beta-cell tumor in culture. Addition of 10 mM NH4C1 to the medium inhibited viral growth by greater than 80% and reduced toxic effects of the virus on cell viability, protein, and DNA synthesis by 30-45%. In addition, synthesis of viral proteins was markedly decreased. Uptake of virus prelabeled with [35S]methionine was not affected by the ammonium; however, cleavage of mu1C, an outer capsid protein of the virus whose cleavage appears to be required for viral replication, was delayed. These results suggest that intracellular processing of reovirus is dependent on a lysosomal pathway and that disruption of this pathway can alter the course of viral infection.
Full text
PDF![1003](https://cdn.ncbi.nlm.nih.gov/pmc/blobs/03ac/423744/87e42455b60c/jcinvest00109-0155.png)
![1004](https://cdn.ncbi.nlm.nih.gov/pmc/blobs/03ac/423744/b8301fa95556/jcinvest00109-0156.png)
![1005](https://cdn.ncbi.nlm.nih.gov/pmc/blobs/03ac/423744/53fc7dd1b23a/jcinvest00109-0157.png)
![1006](https://cdn.ncbi.nlm.nih.gov/pmc/blobs/03ac/423744/baa4bc285e93/jcinvest00109-0158.png)
![1007](https://cdn.ncbi.nlm.nih.gov/pmc/blobs/03ac/423744/f6b49ad43d82/jcinvest00109-0159.png)
Images in this article
Selected References
These references are in PubMed. This may not be the complete list of references from this article.
- Ashwell G., Harford J. Carbohydrate-specific receptors of the liver. Annu Rev Biochem. 1982;51:531–554. doi: 10.1146/annurev.bi.51.070182.002531. [DOI] [PubMed] [Google Scholar]
- Borsa J., Graham A. F. Reovirus: RNA polymerase activity in purified virions. Biochem Biophys Res Commun. 1968 Dec 30;33(6):895–901. doi: 10.1016/0006-291x(68)90396-3. [DOI] [PubMed] [Google Scholar]
- Braunwald J., Nonnenmacher H., Tripier-Darcy F. Ultrastructural and biochemical study of frog virus 3 uptake by BHK-21 cells. J Gen Virol. 1985 Feb;66(Pt 2):283–293. doi: 10.1099/0022-1317-66-2-283. [DOI] [PubMed] [Google Scholar]
- Brown M. S., Kovanen P. T., Goldstein J. L. Regulation of plasma cholesterol by lipoprotein receptors. Science. 1981 May 8;212(4495):628–635. doi: 10.1126/science.6261329. [DOI] [PubMed] [Google Scholar]
- Canning W. M., Fields B. N. Ammonium chloride prevents lytic growth of reovirus and helps to establish persistent infection in mouse L cells. Science. 1983 Feb 25;219(4587):987–988. doi: 10.1126/science.6297010. [DOI] [PubMed] [Google Scholar]
- Carrillo E. C., Giachetti C., Campos R. H. Effect of lysosomotropic agents on the foot-and-mouth disease virus replication. Virology. 1984 Jun;135(2):542–545. doi: 10.1016/0042-6822(84)90208-3. [DOI] [PubMed] [Google Scholar]
- Chang C. T., Zweerink H. J. Fate of parental reovirus in infected cell. Virology. 1971 Dec;46(3):544–555. doi: 10.1016/0042-6822(71)90058-4. [DOI] [PubMed] [Google Scholar]
- Drayna D., Fields B. N. Activation and characterization of the reovirus transcriptase: genetic analysis. J Virol. 1982 Jan;41(1):110–118. doi: 10.1128/jvi.41.1.110-118.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Gazdar A. F., Chick W. L., Oie H. K., Sims H. L., King D. L., Weir G. C., Lauris V. Continuous, clonal, insulin- and somatostatin-secreting cell lines established from a transplantable rat islet cell tumor. Proc Natl Acad Sci U S A. 1980 Jun;77(6):3519–3523. doi: 10.1073/pnas.77.6.3519. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Haspel M. V., Onodera T., Prabhakar B. S., Horita M., Suzuki H., Notkins A. L. Virus-induced autoimmunity: monoclonal antibodies that react with endocrine tissues. Science. 1983 Apr 15;220(4594):304–306. doi: 10.1126/science.6301002. [DOI] [PubMed] [Google Scholar]
- Helenius A., Kartenbeck J., Simons K., Fries E. On the entry of Semliki forest virus into BHK-21 cells. J Cell Biol. 1980 Feb;84(2):404–420. doi: 10.1083/jcb.84.2.404. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Helenius A., Marsh M., White J. Inhibition of Semliki forest virus penetration by lysosomotropic weak bases. J Gen Virol. 1982 Jan;58(Pt 1):47–61. doi: 10.1099/0022-1317-58-1-47. [DOI] [PubMed] [Google Scholar]
- Joklik W. K. Studies on the effect of chymotrypsin on reovirions. Virology. 1972 Sep;49(3):700–715. doi: 10.1016/0042-6822(72)90527-2. [DOI] [PubMed] [Google Scholar]
- 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]
- Maratos-Flier E., Goodman M. J., Fields B. N., Kahn C. R. Differential effects of viral infection on islet and pituitary cell lines. Endocrinology. 1985 Jun;116(6):2430–2437. doi: 10.1210/endo-116-6-2430. [DOI] [PubMed] [Google Scholar]
- Marsh M., Helenius A. Adsorptive endocytosis of Semliki Forest virus. J Mol Biol. 1980 Sep 25;142(3):439–454. doi: 10.1016/0022-2836(80)90281-8. [DOI] [PubMed] [Google Scholar]
- Maxfield F. R., Schlessinger J., Shechter Y., Pastan I., Willingham M. C. Collection of insulin, EGF and alpha2-macroglobulin in the same patches on the surface of cultured fibroblasts and common internalization. Cell. 1978 Aug;14(4):805–810. doi: 10.1016/0092-8674(78)90336-7. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Ohkuma S., Poole B. Fluorescence probe measurement of the intralysosomal pH in living cells and the perturbation of pH by various agents. Proc Natl Acad Sci U S A. 1978 Jul;75(7):3327–3331. doi: 10.1073/pnas.75.7.3327. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Onodera T., Jenson A. B., Yoon J. W., Notkins A. L. Virus-induced diabetes mellitus: reovirus infection of pancreatic beta cells in mice. Science. 1978 Aug 11;201(4355):529–531. doi: 10.1126/science.208156. [DOI] [PubMed] [Google Scholar]
- Onodera T., Toniolo A., Ray U. R., Jenson A. B., Knazek R. A., Notkins A. L. Virus-induced diabetes mellitus. XX. Polyendocrinopathy and autoimmunity. J Exp Med. 1981 Jun 1;153(6):1457–1473. doi: 10.1084/jem.153.6.1457. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Praz G. A., Halban P. A., Wollheim C. B., Blondel B., Strauss A. J., Renold A. E. Regulation of immunoreactive-insulin release from a rat cell line (RINm5F). Biochem J. 1983 Feb 15;210(2):345–352. doi: 10.1042/bj2100345. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Ramig R. F., Cross R. K., Fields B. N. Genome RNAs and polypeptides of reovirus serotypes 1, 2, and 3. J Virol. 1977 Jun;22(3):726–733. doi: 10.1128/jvi.22.3.726-733.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Sandvig K., Olsnes S. Diphtheria toxin entry into cells is facilitated by low pH. J Cell Biol. 1980 Dec;87(3 Pt 1):828–832. doi: 10.1083/jcb.87.3.828. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 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]
- Shatkin A. J., LaFiandra A. J. Transcription by infectious subviral particles of reovirus. J Virol. 1972 Oct;10(4):698–706. doi: 10.1128/jvi.10.4.698-706.1972. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Shatkin A. J., Sipe J. D. RNA polymerase activity in purified reoviruses. Proc Natl Acad Sci U S A. 1968 Dec;61(4):1462–1469. doi: 10.1073/pnas.61.4.1462. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 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]
- 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]
- Yamashiro D. J., Maxfield F. R. Acidification of endocytic compartments and the intracellular pathways of ligands and receptors. J Cell Biochem. 1984;26(4):231–246. doi: 10.1002/jcb.240260404. [DOI] [PubMed] [Google Scholar]
- Yoon J. W., Selvaggio S., Onodera T., Wheeler J., Jenson A. B. Infection of cultured human pancreatic B cells with reovirus type 3. Diabetologia. 1981 Apr;20(4):462–467. doi: 10.1007/BF00253408. [DOI] [PubMed] [Google Scholar]
- Yoshimura A., Ohnishi S. Uncoating of influenza virus in endosomes. J Virol. 1984 Aug;51(2):497–504. doi: 10.1128/jvi.51.2.497-504.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Zeichhardt H., Wetz K., Willingmann P., Habermehl K. O. Entry of poliovirus type 1 and Mouse Elberfeld (ME) virus into HEp-2 cells: receptor-mediated endocytosis and endosomal or lysosomal uncoating. J Gen Virol. 1985 Mar;66(Pt 3):483–492. doi: 10.1099/0022-1317-66-3-483. [DOI] [PubMed] [Google Scholar]