Skip to main content
Journal of Virology logoLink to Journal of Virology
. 1981 Jul;39(1):263–272. doi: 10.1128/jvi.39.1.263-272.1981

Subcellular localization of the env-related glycoproteins in Friend erythroleukemia cells.

D S Lyles, K A McConnell
PMCID: PMC171285  PMID: 6944507

Abstract

A scheme was developed for the subcellular fractionation of murine erythroleukemia cells transformed by Friend leukemia virus. The subcellular localization of the env-related glycoproteins was determined by immune precipitation with antiserum against gp70, the envelope glycoprotein of the helper virus, followed by gel electrophoresis. In cells labeled for 2 h with [35S]methionine, the glycoprotein encoded by the defective spleen focus-forming virus, gp55SFFV, was found primarily in the nuclear fraction and in fractions containing dense cytoplasmic membranes such as endoplasmic reticulum. A similar distribution was noted for gp85env, the precursor to gp70. The concentration of viral glycoproteins in the nuclear fraction could not be accounted for by contamination with endoplasmic reticulum. In pulse-chase experiments, neither glycoprotein underwent major redistribution. However, labeled gp85env disappeared from intracellular membranes with a half-time of 30 min to 1 h, whereas labeled gp55SFFV was stable during a 2-h chase. In plasma membrane preparations with very low levels of contamination with endoplasmic reticulum, gp70 was the major viral env-related glycoprotein detected; a minor amount of gp55SFFV and no gp85env could be detected. The unexpected result of these experiments is the amount of viral glycoproteins found in the nuclear fraction. Presence of viral proteins in the nucleus could be relevant to the mechanism of viral leukemogenesis.

Full text

PDF

Images in this article

Selected References

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

  1. Bonner W. M., Laskey R. A. A film detection method for tritium-labelled proteins and nucleic acids in polyacrylamide gels. Eur J Biochem. 1974 Jul 1;46(1):83–88. doi: 10.1111/j.1432-1033.1974.tb03599.x. [DOI] [PubMed] [Google Scholar]
  2. Bosselman R. A., Van Griensven L. J., Vogt M., Verma I. M. Genome organization of retroviruses IX. Analysis of the genomes of Friend spleen focus-forming (F-SFFV) and helper murine leukemia viruses by heteroduplex-formation. Virology. 1980 Apr 15;102(1):234–239. doi: 10.1016/0042-6822(80)90088-4. [DOI] [PubMed] [Google Scholar]
  3. Bowen H. A., Lyles D. S. Structure of Sendai viral proteins in plasma membranes of virus-infected cells. J Virol. 1981 Mar;37(3):1079–1082. doi: 10.1128/jvi.37.3.1079-1082.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Caliguiri L. A., Tamm I. The role of cytoplasmic membranes in poliovirus biosynthesis. Virology. 1970 Sep;42(1):100–111. doi: 10.1016/0042-6822(70)90242-4. [DOI] [PubMed] [Google Scholar]
  5. Cohen C. M., Kalish D. I., Jacobson B. S., Branton D. Membrane isolation on polylysine-coated beads. Plasma membrane from HeLa cells. J Cell Biol. 1977 Oct;75(1):119–134. doi: 10.1083/jcb.75.1.119. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Dresler S., Ruta M., Murray M. J., Kabat D. Glycoprotein encoded by the Friend spleen focus-forming virus. J Virol. 1979 May;30(2):564–575. doi: 10.1128/jvi.30.2.564-575.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Evans L. H., Dresler S., Kabat D. Synthesis and glycosylation of polyprotein precursors to the internal core proteins of Friend murine leukemia virus. J Virol. 1977 Dec;24(3):865–874. doi: 10.1128/jvi.24.3.865-874.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Hay A. J. Studies on the formation of the influenza virus envelope. Virology. 1974 Aug;60(2):398–418. doi: 10.1016/0042-6822(74)90335-3. [DOI] [PubMed] [Google Scholar]
  9. Kabat D., Ruta M., Murray M. J., Polonoff E. Immunoselection of mutants deficient in cell surface glycoproteins encoded by murine erythroleukemia viruses. Proc Natl Acad Sci U S A. 1980 Jan;77(1):57–61. doi: 10.1073/pnas.77.1.57. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Kay R. R., Johnston I. R. Rapid isolation of nuclear envelopes from rat liver. Methods Cell Biol. 1977;15:277–287. doi: 10.1016/s0091-679x(08)60220-4. [DOI] [PubMed] [Google Scholar]
  11. 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]
  12. Krueger J. G., Wang E., Garber E. A., Goldberg A. R. Differences in intracellular location of pp60src in rat and chicken cells transformed by Rous sarcoma virus. Proc Natl Acad Sci U S A. 1980 Jul;77(7):4142–4146. doi: 10.1073/pnas.77.7.4142. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. LOWRY O. H., ROSEBROUGH N. J., FARR A. L., RANDALL R. J. Protein measurement with the Folin phenol reagent. J Biol Chem. 1951 Nov;193(1):265–275. [PubMed] [Google Scholar]
  14. 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]
  15. Lamb R. A., Choppin P. W. The synthesis of Sendai virus polypeptides in infected cells. II. Intracellular distribution of polypeptides. Virology. 1977 Sep;81(2):371–381. doi: 10.1016/0042-6822(77)90153-2. [DOI] [PubMed] [Google Scholar]
  16. Lenard J. Virus envelopes and plasma membranes. Annu Rev Biophys Bioeng. 1978;7:139–165. doi: 10.1146/annurev.bb.07.060178.001035. [DOI] [PubMed] [Google Scholar]
  17. Markwell M. A., Fox C. F. Surface-specific iodination of membrane proteins of viruses and eucaryotic cells using 1,3,4,6-tetrachloro-3alpha,6alpha-diphenylglycoluril. Biochemistry. 1978 Oct 31;17(22):4807–4817. doi: 10.1021/bi00615a031. [DOI] [PubMed] [Google Scholar]
  18. Montelaro R. C., Bolognesi D. P. Structure and morphogenesis of type-C retroviruses. Adv Cancer Res. 1978;28:63–89. doi: 10.1016/s0065-230x(08)60646-6. [DOI] [PubMed] [Google Scholar]
  19. Racevskis J., Koch G. Synthesis and processing of viral proteins in Friend erythroleukemia cell lines. Virology. 1978 Jun 15;87(2):354–365. doi: 10.1016/0042-6822(78)90140-x. [DOI] [PubMed] [Google Scholar]
  20. Racevskis J., Koch G. Viral protein synthesis in Friend erythroleukemia cell lines. J Virol. 1977 Jan;21(1):328–337. doi: 10.1128/jvi.21.1.328-337.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Rothman J. E., Fine R. E. Coated vesicles transport newly synthesized membrane glycoproteins from endoplasmic reticulum to plasma membrane in two successive stages. Proc Natl Acad Sci U S A. 1980 Feb;77(2):780–784. doi: 10.1073/pnas.77.2.780. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Ruscetti S. K., Linemeyer D., Feild J., Troxler D., Scolnick E. M. Characterization of a protein found in cells infected with the spleen focus-forming virus that shares immunological cross-reactivity with the gp70 found in mink cell focus-inducing virus particles. J Virol. 1979 Jun;30(3):787–798. doi: 10.1128/jvi.30.3.787-798.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Ruscetti S., Troxler D., Linemeyer D., Scolnick E. Three laboratory strains of spleen focus-forming virus: comparison of their genomes and translational products. J Virol. 1980 Jan;33(1):140–151. doi: 10.1128/jvi.33.1.140-151.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Ruta M., Kabat D. Plasma membrane glycoproteins encoded by cloned Rauscher and Friend spleen focus-forming viruses. J Virol. 1980 Sep;35(3):844–853. doi: 10.1128/jvi.35.3.844-853.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Schultz A. M., Ruscetti S. K., Scolnick E. M., Oroszlan S. The env-gene of the spleen focus-forming virus lacks expression of p15(E) determinants. Virology. 1980 Dec;107(2):537–542. doi: 10.1016/0042-6822(80)90322-0. [DOI] [PubMed] [Google Scholar]
  26. Sottocasa G. L., Kuylenstierna B., Ernster L., Bergstrand A. An electron-transport system associated with the outer membrane of liver mitochondria. A biochemical and morphological study. J Cell Biol. 1967 Feb;32(2):415–438. doi: 10.1083/jcb.32.2.415. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Sutcliffe J. G., Shinnick T. M., Green N., Liu F. T., Niman H. L., Lerner R. A. Chemical synthesis of a polypeptide predicted from nucleotide sequence allows detection of a new retroviral gene product. Nature. 1980 Oct 30;287(5785):801–805. doi: 10.1038/287801a0. [DOI] [PubMed] [Google Scholar]
  28. Troxler D. H., Lowy D., Howk R., Young H., Scolnick E. M. Friend strain of spleen focus-forming virus is a recombinant between ecotropic murine type C virus and the env gene region of xenotropic type C virus. Proc Natl Acad Sci U S A. 1977 Oct;74(10):4671–4675. doi: 10.1073/pnas.74.10.4671. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Troxler D. H., Ruscetti S. K., Scolnick E. M. The molecular biology of Friend virus. Biochim Biophys Acta. 1980 Sep 22;605(3):305–324. doi: 10.1016/0304-419x(80)90014-1. [DOI] [PubMed] [Google Scholar]
  30. Witte O. N., Tsukamoto-Adey A., Weissman I. L. Cellular maturation of oncornavirus glycoproteins: topological arrangement of precursor and product forms in cellular membranes. Virology. 1977 Feb;76(2):539–553. doi: 10.1016/0042-6822(77)90236-7. [DOI] [PubMed] [Google Scholar]

Articles from Journal of Virology are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES