Abstract
Using morphological and cell biological techniques, we have shown that the RNA replicase of Semliki Forest and Sindbis virus (two closely related alphaviruses) is located in complex ribonucleoprotein structures associated with the cytoplasmic surface of modified secondary lysosomes and endosomes. These nucleoprotein complexes often form a bridge between the membrane of the endocytic vacuole and the rough endoplasmic reticulum where the synthesis of the structural proteins of these viruses occurs. The results suggest that these cytopathic vacuoles constitute sites not only for viral RNA synthesis, but also for translation of structural proteins, and for the assembly of nucleocapsids.
Full Text
The Full Text of this article is available as a PDF (4.2 MB).
Selected References
These references are in PubMed. This may not be the complete list of references from this article.
- Acheson N. H., Tamm I. Replication of Semliki Forest virus: an electron microscopic study. Virology. 1967 May;32(1):128–143. doi: 10.1016/0042-6822(67)90261-9. [DOI] [PubMed] [Google Scholar]
- Balch W. E., Rothman J. E. Characterization of protein transport between successive compartments of the Golgi apparatus: asymmetric properties of donor and acceptor activities in a cell-free system. Arch Biochem Biophys. 1985 Jul;240(1):413–425. doi: 10.1016/0003-9861(85)90046-3. [DOI] [PubMed] [Google Scholar]
- Beckers C. J., Keller D. S., Balch W. E. Semi-intact cells permeable to macromolecules: use in reconstitution of protein transport from the endoplasmic reticulum to the Golgi complex. Cell. 1987 Aug 14;50(4):523–534. doi: 10.1016/0092-8674(87)90025-0. [DOI] [PubMed] [Google Scholar]
- Bradford M. M. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem. 1976 May 7;72:248–254. doi: 10.1006/abio.1976.9999. [DOI] [PubMed] [Google Scholar]
- Brown W. J., Farquhar M. G. Accumulation of coated vesicles bearing mannose 6-phosphate receptors for lysosomal enzymes in the Golgi region of I-cell fibroblasts. Proc Natl Acad Sci U S A. 1984 Aug;81(16):5135–5139. doi: 10.1073/pnas.81.16.5135. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Chamberlain J. P. Fluorographic detection of radioactivity in polyacrylamide gels with the water-soluble fluor, sodium salicylate. Anal Biochem. 1979 Sep 15;98(1):132–135. doi: 10.1016/0003-2697(79)90716-4. [DOI] [PubMed] [Google Scholar]
- Clewley J. P., Kennedy S. I. Purification and polypeptide composition of Semliki Forest virus RNA polymerase. J Gen Virol. 1976 Sep;32(3):395–411. doi: 10.1099/0022-1317-32-3-395. [DOI] [PubMed] [Google Scholar]
- Dunn W. A., Hubbard A. L., Aronson N. N., Jr Low temperature selectively inhibits fusion between pinocytic vesicles and lysosomes during heterophagy of 125I-asialofetuin by the perfused rat liver. J Biol Chem. 1980 Jun 25;255(12):5971–5978. [PubMed] [Google Scholar]
- Friedman R. M., Levin J. G., Grimley P. M., Berezesky I. K. Membrane-associated replication complex in arbovirus infection. J Virol. 1972 Sep;10(3):504–515. doi: 10.1128/jvi.10.3.504-515.1972. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Fuller S. D. The T=4 envelope of Sindbis virus is organized by interactions with a complementary T=3 capsid. Cell. 1987 Mar 27;48(6):923–934. doi: 10.1016/0092-8674(87)90701-x. [DOI] [PubMed] [Google Scholar]
- Gomatos P. J., Käriäinen L., Keränen S., Ranki M., Sawicki D. L. Semliki Forest virus replication complex capable of synthesizing 42S and 26S nascent RNA chains. J Gen Virol. 1980 Jul;49(1):61–69. doi: 10.1099/0022-1317-49-1-61. [DOI] [PubMed] [Google Scholar]
- Green S. A., Zimmer K. P., Griffiths G., Mellman I. Kinetics of intracellular transport and sorting of lysosomal membrane and plasma membrane proteins. J Cell Biol. 1987 Sep;105(3):1227–1240. doi: 10.1083/jcb.105.3.1227. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Grimley P. M., Berezesky I. K., Friedman R. M. Cytoplasmic structures associated with an arbovirus infection: loci of viral ribonucleic acid synthesis. J Virol. 1968 Nov;2(11):1326–1338. doi: 10.1128/jvi.2.11.1326-1338.1968. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Grimley P. M., Levin J. G., Berezesky I. K., Friedman R. M. Specific membranous structures associated with the replication of group A arboviruses. J Virol. 1972 Sep;10(3):492–503. doi: 10.1128/jvi.10.3.492-503.1972. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Hardy W. R., Strauss J. H. Processing the nonstructural polyproteins of Sindbis virus: study of the kinetics in vivo by using monospecific antibodies. J Virol. 1988 Mar;62(3):998–1007. doi: 10.1128/jvi.62.3.998-1007.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Harms E., Kern H., Schneider J. A. Human lysosomes can be purified from diploid skin fibroblasts by free-flow electrophoresis. Proc Natl Acad Sci U S A. 1980 Oct;77(10):6139–6143. doi: 10.1073/pnas.77.10.6139. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Hawley D. M., Tsou K. C., Hodes M. E. Preparation, properties, and uses of two fluorogenic substrates for the detection of 5'-(venom) and 3'-(spleen) nucleotide phosphodiesterases. Anal Biochem. 1981 Oct;117(1):18–23. doi: 10.1016/0003-2697(81)90684-9. [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., Kartenbeck J. The effects of octylglucoside on the Semliki forest virus membrane. Evidence for a spike-protein--nucleocapsid interaction. Eur J Biochem. 1980 May;106(2):613–618. doi: 10.1111/j.1432-1033.1980.tb04609.x. [DOI] [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]
- Keränen S., Ruohonen L. Nonstructural proteins of Semliki Forest virus: synthesis, processing, and stability in infected cells. J Virol. 1983 Sep;47(3):505–515. doi: 10.1128/jvi.47.3.505-515.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Kielian M. C., Keränen S., Käriäinen L., Helenius A. Membrane fusion mutants of Semliki Forest virus. J Cell Biol. 1984 Jan;98(1):139–145. doi: 10.1083/jcb.98.1.139. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Kielian M. C., Marsh M., Helenius A. Kinetics of endosome acidification detected by mutant and wild-type Semliki Forest virus. EMBO J. 1986 Dec 1;5(12):3103–3109. doi: 10.1002/j.1460-2075.1986.tb04616.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Käriäinen L., Simons K., von Bonsdorff C. H. Studies in subviral components of Semliki Forest virus. Ann Med Exp Biol Fenn. 1969;47(4):235–248. [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]
- Lewis V., Green S. A., Marsh M., Vihko P., Helenius A., Mellman I. Glycoproteins of the lysosomal membrane. J Cell Biol. 1985 Jun;100(6):1839–1847. doi: 10.1083/jcb.100.6.1839. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Lopez S., Bell J. R., Strauss E. G., Strauss J. H. The nonstructural proteins of Sindbis virus as studied with an antibody specific for the C terminus of the nonstructural readthrough polyprotein. Virology. 1985 Mar;141(2):235–247. doi: 10.1016/0042-6822(85)90254-5. [DOI] [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]
- Marsh M., Griffiths G., Dean G. E., Mellman I., Helenius A. Three-dimensional structure of endosomes in BHK-21 cells. Proc Natl Acad Sci U S A. 1986 May;83(9):2899–2903. doi: 10.1073/pnas.83.9.2899. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Marsh M., Schmid S., Kern H., Harms E., Male P., Mellman I., Helenius A. Rapid analytical and preparative isolation of functional endosomes by free flow electrophoresis. J Cell Biol. 1987 Apr;104(4):875–886. doi: 10.1083/jcb.104.4.875. [DOI] [PMC free article] [PubMed] [Google Scholar]
- McLean I. W., Nakane P. K. Periodate-lysine-paraformaldehyde fixative. A new fixation for immunoelectron microscopy. J Histochem Cytochem. 1974 Dec;22(12):1077–1083. doi: 10.1177/22.12.1077. [DOI] [PubMed] [Google Scholar]
- Nilsson J., Ksiazek T., Thyberg J. Endocytosis of cationic and anionic proteins in cultivated arterial smooth muscle cells. Exp Cell Res. 1983 Feb;143(2):359–365. doi: 10.1016/0014-4827(83)90062-9. [DOI] [PubMed] [Google Scholar]
- Pool R. R., Jr, Maurey K. M., Storrie B. Characterization of pinocytic vesicles from CHO cells: resolution of pinosomes from lysosomes by analytical centrifugation. Cell Biol Int Rep. 1983 May;7(5):361–367. doi: 10.1016/0309-1651(83)90076-0. [DOI] [PubMed] [Google Scholar]
- Ranki M., Käriäinen L. Solubilized RNA replication complex from Semliki Forest virus-infected cells. Virology. 1979 Oct 30;98(2):298–307. doi: 10.1016/0042-6822(79)90553-1. [DOI] [PubMed] [Google Scholar]
- Sawicki D. L., Sawicki S. G. Short-lived minus-strand polymerase for Semliki Forest virus. J Virol. 1980 Apr;34(1):108–118. doi: 10.1128/jvi.34.1.108-118.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Scheele C. M., Pfefferkorn E. R. Kinetics of incorporation of structural proteins into Sindbis virions. J Virol. 1969 Apr;3(4):369–375. doi: 10.1128/jvi.3.4.369-375.1969. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Schmid S. L., Fuchs R., Male P., Mellman I. Two distinct subpopulations of endosomes involved in membrane recycling and transport to lysosomes. Cell. 1988 Jan 15;52(1):73–83. doi: 10.1016/0092-8674(88)90532-6. [DOI] [PubMed] [Google Scholar]
- Simmons D. T., Strauss J. H. Replication of Sindbis virus. II. Multiple forms of double-stranded RNA isolated from infected cells. J Mol Biol. 1972 Nov 28;71(3):615–631. doi: 10.1016/s0022-2836(72)80027-5. [DOI] [PubMed] [Google Scholar]
- Simons K., Garoff H. The budding mechanisms of enveloped animal viruses. J Gen Virol. 1980 Sep;50(1):1–21. doi: 10.1099/0022-1317-50-1-1. [DOI] [PubMed] [Google Scholar]
- Simons K., Virta H. Perforated MDCK cells support intracellular transport. EMBO J. 1987 Aug;6(8):2241–2247. doi: 10.1002/j.1460-2075.1987.tb02496.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Strauss E. G., Rice C. M., Strauss J. H. Sequence coding for the alphavirus nonstructural proteins is interrupted by an opal termination codon. Proc Natl Acad Sci U S A. 1983 Sep;80(17):5271–5275. doi: 10.1073/pnas.80.17.5271. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Ukkonen P., Lewis V., Marsh M., Helenius A., Mellman I. Transport of macrophage Fc receptors and Fc receptor-bound ligands to lysosomes. J Exp Med. 1986 Apr 1;163(4):952–971. doi: 10.1084/jem.163.4.952. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Weibel E. R. Stereological principles for morphometry in electron microscopic cytology. Int Rev Cytol. 1969;26:235–302. doi: 10.1016/s0074-7696(08)61637-x. [DOI] [PubMed] [Google Scholar]
- Wengler G., Boege U., Wengler G., Bischoff H., Wahn K. The core protein of the alphavirus Sindbis virus assembles into core-like nucleoproteins with the viral genome RNA and with other single-stranded nucleic acids in vitro. Virology. 1982 Apr 30;118(2):401–410. doi: 10.1016/0042-6822(82)90359-2. [DOI] [PubMed] [Google Scholar]