Skip to main content
NCBI home page
The Journal of Cell Biology logoLink to The Journal of Cell Biology
. 1992 Jun 1;117(5):987–995. doi: 10.1083/jcb.117.5.987

Local expression and exocytosis of viral glycoproteins in multinucleated muscle cells

PMCID: PMC2289478  PMID: 1315787

Abstract

We have analyzed the distribution of enveloped viral infections in multinucleated L6 muscle cells. A temperature-sensitive vesicular stomatitis virus (mutant VSV ts045) was utilized at the nonpermissive temperature (39 degrees C). As expected, the glycoprotein (G protein) of this mutant was restricted to the ER when the multinucleated cells were maintained at 39 degrees C. We demonstrate that this G protein remained localized when the infection was performed at low dose. By 4 h after infection the G protein patches spanned an average of 220 microns. The localization was independent of nuclear positions, showing that the ER was a peripheric structure. Thus, the infection did not recognize nuclear domains characteristic of nuclearly encoded proteins. After release of the 39 degrees C block, transport through a perinuclear compartment into a restricted surface domain lying above the internal G protein patch occurred. Accordingly, the transport pathway was locally restricted. After a 16-h infection the G protein spanned 420 microns, while the matrix protein occupied 700-800 microns of the myotube length. Double infection of multinucleated L6 muscle cells with Semliki Forest virus and VSV at high multiplicities showed that the glycoprotein of each virus occupied intracellular domains which were devoid of the other respective glycoprotein. Taken together, these findings indicate that the viral glycoproteins did not range far from their site of synthesis within the ER or other intracellular membrane compartments in these large cells. This result also suggests that relocation of viral RNA synthesis occurred slowly.

Full Text

The Full Text of this article is available as a PDF (4.0 MB).

Selected References

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

  1. Baron M. D., Garoff H. Mannosidase II and the 135-kDa Golgi-specific antigen recognized monoclonal antibody 53FC3 are the same dimeric protein. J Biol Chem. 1990 Nov 15;265(32):19928–19931. [PubMed] [Google Scholar]
  2. Blondel D., Harmison G. G., Schubert M. Role of matrix protein in cytopathogenesis of vesicular stomatitis virus. J Virol. 1990 Apr;64(4):1716–1725. doi: 10.1128/jvi.64.4.1716-1725.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Burke B., Griffiths G., Reggio H., Louvard D., Warren G. A monoclonal antibody against a 135-K Golgi membrane protein. EMBO J. 1982;1(12):1621–1628. doi: 10.1002/j.1460-2075.1982.tb01364.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Eidelman O., Schlegel R., Tralka T. S., Blumenthal R. pH-dependent fusion induced by vesicular stomatitis virus glycoprotein reconstituted into phospholipid vesicles. J Biol Chem. 1984 Apr 10;259(7):4622–4628. [PubMed] [Google Scholar]
  5. Featherstone C., Griffiths G., Warren G. Newly synthesized G protein of vesicular stomatitis virus is not transported to the Golgi complex in mitotic cells. J Cell Biol. 1985 Dec;101(6):2036–2046. doi: 10.1083/jcb.101.6.2036. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Fontaine B., Changeux J. P. Localization of nicotinic acetylcholine receptor alpha-subunit transcripts during myogenesis and motor endplate development in the chick. J Cell Biol. 1989 Mar;108(3):1025–1037. doi: 10.1083/jcb.108.3.1025. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Froshauer S., Kartenbeck J., Helenius A. Alphavirus RNA replicase is located on the cytoplasmic surface of endosomes and lysosomes. J Cell Biol. 1988 Dec;107(6 Pt 1):2075–2086. doi: 10.1083/jcb.107.6.2075. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Galfrè G., Milstein C. Preparation of monoclonal antibodies: strategies and procedures. Methods Enzymol. 1981;73(Pt B):3–46. doi: 10.1016/0076-6879(81)73054-4. [DOI] [PubMed] [Google Scholar]
  9. Griffiths G., Pfeiffer S., Simons K., Matlin K. Exit of newly synthesized membrane proteins from the trans cisterna of the Golgi complex to the plasma membrane. J Cell Biol. 1985 Sep;101(3):949–964. doi: 10.1083/jcb.101.3.949. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Griffiths G., Simons K. The trans Golgi network: sorting at the exit site of the Golgi complex. Science. 1986 Oct 24;234(4775):438–443. doi: 10.1126/science.2945253. [DOI] [PubMed] [Google Scholar]
  11. Gu Y., Ralston E., Murphy-Erdosh C., Black R. A., Hall Z. W. Acetylcholine receptor in a C2 muscle cell variant is retained in the endoplasmic reticulum. J Cell Biol. 1989 Aug;109(2):729–738. doi: 10.1083/jcb.109.2.729. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Hall Z. W., Ralston E. Nuclear domains in muscle cells. Cell. 1989 Dec 1;59(5):771–772. doi: 10.1016/0092-8674(89)90597-7. [DOI] [PubMed] [Google Scholar]
  13. Knipe D. M., Baltimore D., Lodish H. F. Separate pathways of maturation of the major structural proteins of vesicular stomatitis virus. J Virol. 1977 Mar;21(3):1128–1139. doi: 10.1128/jvi.21.3.1128-1139.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. 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]
  15. 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]
  16. Louvard D. Apical membrane aminopeptidase appears at site of cell-cell contact in cultured kidney epithelial cells. Proc Natl Acad Sci U S A. 1980 Jul;77(7):4132–4136. doi: 10.1073/pnas.77.7.4132. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Louvard D., Reggio H., Warren G. Antibodies to the Golgi complex and the rough endoplasmic reticulum. J Cell Biol. 1982 Jan;92(1):92–107. doi: 10.1083/jcb.92.1.92. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Matlin K. S., Simons K. Reduced temperature prevents transfer of a membrane glycoprotein to the cell surface but does not prevent terminal glycosylation. Cell. 1983 Aug;34(1):233–243. doi: 10.1016/0092-8674(83)90154-x. [DOI] [PubMed] [Google Scholar]
  19. Matlin K., Bainton D. F., Pesonen M., Louvard D., Genty N., Simons K. Transepithelial transport of a viral membrane glycoprotein implanted into the apical plasma membrane of Madin-Darby canine kidney cells. I. Morphological evidence. J Cell Biol. 1983 Sep;97(3):627–637. doi: 10.1083/jcb.97.3.627. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Merlie J. P., Sanes J. R. Concentration of acetylcholine receptor mRNA in synaptic regions of adult muscle fibres. Nature. 1985 Sep 5;317(6032):66–68. doi: 10.1038/317066a0. [DOI] [PubMed] [Google Scholar]
  21. Metsikkö K., Garoff H. Oligomers of the cytoplasmic domain of the p62/E2 membrane protein of Semliki Forest virus bind to the nucleocapsid in vitro. J Virol. 1990 Oct;64(10):4678–4683. doi: 10.1128/jvi.64.10.4678-4683.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Metsikkö K., Garoff H. Role of heterologous and homologous glycoproteins in phenotypic mixing between Sendai virus and vesicular stomatitis virus. J Virol. 1989 Dec;63(12):5111–5118. doi: 10.1128/jvi.63.12.5111-5118.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Metsikkö K., Simons K. The budding mechanism of spikeless vesicular stomatitis virus particles. EMBO J. 1986 Aug;5(8):1913–1920. doi: 10.1002/j.1460-2075.1986.tb04444.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Metsikkö K., van Meer G., Simons K. Reconstitution of the fusogenic activity of vesicular stomatitis virus. EMBO J. 1986 Dec 20;5(13):3429–3435. doi: 10.1002/j.1460-2075.1986.tb04665.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Mintz B., Baker W. W. Normal mammalian muscle differentiation and gene control of isocitrate dehydrogenase synthesis. Proc Natl Acad Sci U S A. 1967 Aug;58(2):592–598. doi: 10.1073/pnas.58.2.592. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Newcomb W. W., Tobin G. J., McGowan J. J., Brown J. C. In vitro reassembly of vesicular stomatitis virus skeletons. J Virol. 1982 Mar;41(3):1055–1062. doi: 10.1128/jvi.41.3.1055-1062.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Pavlath G. K., Rich K., Webster S. G., Blau H. M. Localization of muscle gene products in nuclear domains. Nature. 1989 Feb 9;337(6207):570–573. doi: 10.1038/337570a0. [DOI] [PubMed] [Google Scholar]
  28. Petri W. A., Jr, Wagner R. R. Reconstitution into liposomes of the glycoprotein of vesicular stomatitis virus by detergent dialysis. J Biol Chem. 1979 Jun 10;254(11):4313–4316. [PubMed] [Google Scholar]
  29. Ralston E., Hall Z. W. Intracellular and surface distribution of a membrane protein (CD8) derived from a single nucleus in multinucleated myotubes. J Cell Biol. 1989 Nov;109(5):2345–2352. doi: 10.1083/jcb.109.5.2345. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Ralston E., Hall Z. W. Transfer of a protein encoded by a single nucleus to nearby nuclei in multinucleated myotubes. Science. 1989 Jun 2;244(4908):1066–1069. doi: 10.1126/science.2543074. [DOI] [PubMed] [Google Scholar]
  31. Risau W., Saumweber H., Symmons P. Monoclonal antibodies against a nuclear membrane protein of Drosophila. Localization by indirect immunofluorescence and detection of antigen using a new protein blotting procedure. Exp Cell Res. 1981 May;133(1):47–54. doi: 10.1016/0014-4827(81)90355-4. [DOI] [PubMed] [Google Scholar]
  32. Rotundo R. L. Nucleus-specific translation and assembly of acetylcholinesterase in multinucleated muscle cells. J Cell Biol. 1990 Mar;110(3):715–719. doi: 10.1083/jcb.110.3.715. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Schnitzer T. J., Dickson C., Weiss R. A. Morphological and biochemical characterization of viral particles produced by the tsO45 mutant of vesicular stomatitis virus at restrictive temperature. J Virol. 1979 Jan;29(1):185–195. doi: 10.1128/jvi.29.1.185-195.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Scullion B. F., Hou Y., Puddington L., Rose J. K., Jacobson K. Effects of mutations in three domains of the vesicular stomatitis viral glycoprotein on its lateral diffusion in the plasma membrane. J Cell Biol. 1987 Jul;105(1):69–75. doi: 10.1083/jcb.105.1.69. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Tassin A. M., Maro B., Bornens M. Fate of microtubule-organizing centers during myogenesis in vitro. J Cell Biol. 1985 Jan;100(1):35–46. doi: 10.1083/jcb.100.1.35. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Tassin A. M., Paintrand M., Berger E. G., Bornens M. The Golgi apparatus remains associated with microtubule organizing centers during myogenesis. J Cell Biol. 1985 Aug;101(2):630–638. doi: 10.1083/jcb.101.2.630. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Yaffe D. Retention of differentiation potentialities during prolonged cultivation of myogenic cells. Proc Natl Acad Sci U S A. 1968 Oct;61(2):477–483. doi: 10.1073/pnas.61.2.477. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. Zilberstein A., Snider M. D., Porter M., Lodish H. F. Mutants of vesicular stomatitis virus blocked at different stages in maturation of the viral glycoprotein. Cell. 1980 Sep;21(2):417–427. doi: 10.1016/0092-8674(80)90478-x. [DOI] [PubMed] [Google Scholar]

Articles from The Journal of Cell Biology are provided here courtesy of The Rockefeller University Press

RESOURCES