Abstract
Lysis of HeLa cells infected with poliovirus revealed intact virus; 135S particles, devoid of VP4 but containing the viral RNA; and 80S empty capsids. During infection the kinetics of poliovirus uncoating showed a continuous decrease of intact virus, while the number of 135S particles and empty shells increased. After 1.5 h of infection conformational transition to altered particles resulted in complete disappearance of intact virions. To investigate the mechanism of poliovirus uncoating, which has been suggested to depend on low pH in endosomal compartments of cells, we used lysosomotropic amines to raise the pH in these vesicles. In the presence of ammonium chloride, however, the kinetics of uncoating were similar to those for untreated cells, whereas in cells treated with methylamine, monensin, or chloroquine, uncoating was merely delayed by about 30 min. This effect could be attributed to a delay of virus entry into cells after treatment with methylamine and monensin, whereas chloroquine stabilized the viral capsid itself. Thus, elevation of endosomal pH did not affect virus uncoating. We therefore propose a mechanism of poliovirus uncoating which is independent of low pH.
Full text
PDFSelected References
These references are in PubMed. This may not be the complete list of references from this article.
- Belsham G. J., Bostock C. J. Studies on the infectivity of foot-and-mouth disease virus RNA using microinjection. J Gen Virol. 1988 Feb;69(Pt 2):265–274. doi: 10.1099/0022-1317-69-2-265. [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]
- Cassell S., Edwards J., Brown D. T. Effects of lysosomotropic weak bases on infection of BHK-21 cells by Sindbis virus. J Virol. 1984 Dec;52(3):857–864. doi: 10.1128/jvi.52.3.857-864.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Chardonnet Y., Dales S. Early events in the interaction of adenoviruses with HeLa cells. I. Penetration of type 5 and intracellular release of the DNA genome. Virology. 1970 Mar;40(3):462–477. doi: 10.1016/0042-6822(70)90189-3. [DOI] [PubMed] [Google Scholar]
- Dautry-Varsat A., Ciechanover A., Lodish H. F. pH and the recycling of transferrin during receptor-mediated endocytosis. Proc Natl Acad Sci U S A. 1983 Apr;80(8):2258–2262. doi: 10.1073/pnas.80.8.2258. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Dickson R. B., Schlegel R., Willingham M. C., Pastan I. H. Reversible and irreversible inhibitors of clustering of alpha 2M in clathrin-coated pits on the surface of fibroblasts. Exp Cell Res. 1982 Jul;140(1):215–225. doi: 10.1016/0014-4827(82)90171-9. [DOI] [PubMed] [Google Scholar]
- Diment S., Leech M. S., Stahl P. D. Cathepsin D is membrane-associated in macrophage endosomes. J Biol Chem. 1988 May 15;263(14):6901–6907. [PubMed] [Google Scholar]
- Diment S., Martin K. J., Stahl P. D. Cleavage of parathyroid hormone in macrophage endosomes illustrates a novel pathway for intracellular processing of proteins. J Biol Chem. 1989 Aug 15;264(23):13403–13406. [PubMed] [Google Scholar]
- Everaert L., Vrijsen R., Boeyé A. Eclipse products of poliovirus after cold-synchronized infection of HeLa cells. Virology. 1989 Jul;171(1):76–82. doi: 10.1016/0042-6822(89)90512-6. [DOI] [PubMed] [Google Scholar]
- FENWICK M. L., COOPER P. D. Early interactions between poliovirus and ERK cells: some observations on the nature and significance of the rejected particles. Virology. 1962 Oct;18:212–223. doi: 10.1016/0042-6822(62)90007-7. [DOI] [PubMed] [Google Scholar]
- Filman D. J., Syed R., Chow M., Macadam A. J., Minor P. D., Hogle J. M. Structural factors that control conformational transitions and serotype specificity in type 3 poliovirus. EMBO J. 1989 May;8(5):1567–1579. doi: 10.1002/j.1460-2075.1989.tb03541.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Galloway C. J., Dean G. E., Marsh M., Rudnick G., Mellman I. Acidification of macrophage and fibroblast endocytic vesicles in vitro. Proc Natl Acad Sci U S A. 1983 Jun;80(11):3334–3338. doi: 10.1073/pnas.80.11.3334. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Guttman N., Baltimore D. A plasma membrane component able to bind and alter virions of poliovirus type 1: studies on cell-free alteration using a simplified assay. Virology. 1977 Oct 1;82(1):25–36. doi: 10.1016/0042-6822(77)90029-0. [DOI] [PubMed] [Google Scholar]
- HOLLAND J. J., HOYER B. H. Early stages of enterovirus infection. Cold Spring Harb Symp Quant Biol. 1962;27:101–112. doi: 10.1101/sqb.1962.027.001.013. [DOI] [PubMed] [Google Scholar]
- Harford J., Bridges K., Ashwell G., Klausner R. D. Intracellular dissociation of receptor-bound asialoglycoproteins in cultured hepatocytes. A pH-mediated nonlysosomal event. J Biol Chem. 1983 Mar 10;258(5):3191–3197. [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]
- 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]
- Lonberg-Holm K., Gosser L. B., Kauer J. C. Early alteration of poliovirus in infected cells and its specific inhibition. J Gen Virol. 1975 Jun;27(3):329–342. doi: 10.1099/0022-1317-27-3-329. [DOI] [PubMed] [Google Scholar]
- Lonberg-Holm K., Whiteley N. M. Physical and metabolic requirements for early interaction of poliovirus and human rhinovirus with HeLa cells. J Virol. 1976 Sep;19(3):857–870. doi: 10.1128/jvi.19.3.857-870.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Madshus I. H., Olsnes S., Sandvig K. Different pH requirements for entry of the two picornaviruses, human rhinovirus 2 and murine encephalomyocarditis virus. Virology. 1984 Dec;139(2):346–357. doi: 10.1016/0042-6822(84)90380-5. [DOI] [PubMed] [Google Scholar]
- Madshus I. H., Olsnes S., Sandvig K. Mechanism of entry into the cytosol of poliovirus type 1: requirement for low pH. J Cell Biol. 1984 Apr;98(4):1194–1200. doi: 10.1083/jcb.98.4.1194. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Marsh M., Bolzau E., Helenius A. Penetration of Semliki Forest virus from acidic prelysosomal vacuoles. Cell. 1983 Mar;32(3):931–940. doi: 10.1016/0092-8674(83)90078-8. [DOI] [PubMed] [Google Scholar]
- Matlin K. S., Reggio H., Helenius A., Simons K. Pathway of vesicular stomatitis virus entry leading to infection. J Mol Biol. 1982 Apr 15;156(3):609–631. doi: 10.1016/0022-2836(82)90269-8. [DOI] [PubMed] [Google Scholar]
- Maxfield F. R. Weak bases and ionophores rapidly and reversibly raise the pH of endocytic vesicles in cultured mouse fibroblasts. J Cell Biol. 1982 Nov;95(2 Pt 1):676–681. doi: 10.1083/jcb.95.2.676. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Mendelsohn C. L., Wimmer E., Racaniello V. R. Cellular receptor for poliovirus: molecular cloning, nucleotide sequence, and expression of a new member of the immunoglobulin superfamily. Cell. 1989 Mar 10;56(5):855–865. doi: 10.1016/0092-8674(89)90690-9. [DOI] [PubMed] [Google Scholar]
- Neubauer C., Frasel L., Kuechler E., Blaas D. Mechanism of entry of human rhinovirus 2 into HeLa cells. Virology. 1987 May;158(1):255–258. doi: 10.1016/0042-6822(87)90264-9. [DOI] [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]
- Poole B., Ohkuma S. Effect of weak bases on the intralysosomal pH in mouse peritoneal macrophages. J Cell Biol. 1981 Sep;90(3):665–669. doi: 10.1083/jcb.90.3.665. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Schlegel R., Willingham M., Pastan I. Monensin blocks endocytosis of vesicular stomatitis virus. Biochem Biophys Res Commun. 1981 Oct 15;102(3):992–998. doi: 10.1016/0006-291x(81)91636-3. [DOI] [PubMed] [Google Scholar]
- Seglen P. O. Inhibitors of lysosomal function. Methods Enzymol. 1983;96:737–764. doi: 10.1016/s0076-6879(83)96063-9. [DOI] [PubMed] [Google Scholar]
- Smith T. J., Kremer M. J., Luo M., Vriend G., Arnold E., Kamer G., Rossmann M. G., McKinlay M. A., Diana G. D., Otto M. J. The site of attachment in human rhinovirus 14 for antiviral agents that inhibit uncoating. Science. 1986 Sep 19;233(4770):1286–1293. doi: 10.1126/science.3018924. [DOI] [PubMed] [Google Scholar]
- Sturzenbecker L. J., Nibert M., Furlong D., Fields B. N. Intracellular digestion of reovirus particles requires a low pH and is an essential step in the viral infectious cycle. J Virol. 1987 Aug;61(8):2351–2361. doi: 10.1128/jvi.61.8.2351-2361.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Tycko B., Maxfield F. R. Rapid acidification of endocytic vesicles containing alpha 2-macroglobulin. Cell. 1982 Mar;28(3):643–651. doi: 10.1016/0092-8674(82)90219-7. [DOI] [PubMed] [Google Scholar]
- Wetz K., Willingmann P., Zeichhardt H., Habermehl K. O. Neutralization of poliovirus by polyclonal antibodies requires binding of a single IgG molecule per virion. Arch Virol. 1986;91(3-4):207–220. doi: 10.1007/BF01314281. [DOI] [PubMed] [Google Scholar]
- Wetz K., Zeichhardt H., Willingmann P., Habermehl K. O. Dense particles and slow sedimenting particles produced by ultraviolet irradiation of poliovirus. J Gen Virol. 1983 Jun;64(Pt 6):1263–1275. doi: 10.1099/0022-1317-64-6-1263. [DOI] [PubMed] [Google Scholar]
- Wibo M., Poole B. Protein degradation in cultured cells. II. The uptake of chloroquine by rat fibroblasts and the inhibition of cellular protein degradation and cathepsin B1. J Cell Biol. 1974 Nov;63(2 Pt 1):430–440. doi: 10.1083/jcb.63.2.430. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Wileman T., Harding C., Stahl P. Receptor-mediated endocytosis. Biochem J. 1985 Nov 15;232(1):1–14. doi: 10.1042/bj2320001. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Willingmann P., Barnert H., Zeichhardt H., Habermehl K. O. Recovery of structurally intact and infectious poliovirus type 1 from HeLa cells during receptor-mediated endocytosis. Virology. 1989 Feb;168(2):417–420. doi: 10.1016/0042-6822(89)90286-9. [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]
- Zajac I., Crowell R. L. Differential inhibition of attachment and eclipse activities of HeLa cells for enteroviruses. J Virol. 1969 Apr;3(4):422–428. doi: 10.1128/jvi.3.4.422-428.1969. [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]
- de Duve C. Lysosomes revisited. Eur J Biochem. 1983 Dec 15;137(3):391–397. doi: 10.1111/j.1432-1033.1983.tb07841.x. [DOI] [PubMed] [Google Scholar]