Skip to main content
PMC home page
Journal of Virology logoLink to Journal of Virology
. 1991 Mar;65(3):1114–1123. doi: 10.1128/jvi.65.3.1114-1123.1991

Processing of the glycoprotein of feline immunodeficiency virus: effect of inhibitors of glycosylation.

E B Stephens 1, E Monck 1, K Reppas 1, E J Butfiloski 1
PMCID: PMC239877  PMID: 1847441

Abstract

The processing and transport of the envelope glycoprotein complex of feline immunodeficiency virus (FIV) in the persistently infected Crandell feline kidney (CRFK) cell line were investigated. Pulse-chase analyses revealed that the glycoprotein is synthesized as a precursor with an Mr of 145,000 (gp145) and is quickly trimmed to a molecule with an Mr of 130,000 (gp130). Treatment of gp130 with endoglycosidase H (endo H) resulted in a protein with an Mr of 75,000, indicating that nearly half the weight of the gp130 precursor consists of endo H-sensitive glycans during biosynthesis. Chase periods of up to 8 h revealed intermediates during the further processing of this glycoprotein precursor. Initially, two minor protein species with apparent Mrs of 100,000 and 90,000 were detected along with gp130. At later chase times these two species appeared to migrate as a single dominant species with an Mr of 95,000 (gp95). Concomitant with the appearance of gp95 was another protein with an Mr of approximately 40,000 (gp40). Chase periods of up to 8 h revealed that approximately half of the precursor was processed into the gp95-gp40 complex within 4 h. gp95 was efficiently transported from the cell into the culture medium by 1 to 2 h after labeling, whereas gp40 was not observed to be released from infected CRFK cells. Analysis of the processing in the presence of monensin, castanospermine, and swainsonine also suggests the existence of these intermediates in the processing of this lentivirus glycoprotein. As with human immunodeficiency virus, virus produced in the presence of glucosidase inhibitors and reduced infectivity for T-lymphocyte cultures.

Full text

PDF
1114

Images in this article

1116Image
on p.1116
1116Image
on p.1116
1117Image
on p.1117
1117Image
on p.1117
1117Image
on p.1117
1118Image
on p.1118
1119Image
on p.1119
1120Image
on p.1120

Selected References

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

  1. Blough H. A., Pauwels R., De Clercq E., Cogniaux J., Sprecher-Goldberger S., Thiry L. Glycosylation inhibitors block the expression of LAV/HTLV-III (HIV) glycoproteins. Biochem Biophys Res Commun. 1986 Nov 26;141(1):33–38. doi: 10.1016/s0006-291x(86)80330-8. [DOI] [PubMed] [Google Scholar]
  2. Brunner D., Pedersen N. C. Infection of peritoneal macrophages in vitro and in vivo with feline immunodeficiency virus. J Virol. 1989 Dec;63(12):5483–5488. doi: 10.1128/jvi.63.12.5483-5488.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Collins J. K., Britt W. J., Chesebro B. Cytotoxic T lymphocyte recognition of gp70 on Friend virus-induced erythroleukemia cell clones. J Immunol. 1980 Sep;125(3):1318–1324. [PubMed] [Google Scholar]
  4. Dewar R. L., Vasudevachari M. B., Natarajan V., Salzman N. P. Biosynthesis and processing of human immunodeficiency virus type 1 envelope glycoproteins: effects of monensin on glycosylation and transport. J Virol. 1989 Jun;63(6):2452–2456. doi: 10.1128/jvi.63.6.2452-2456.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Dow S. W., Poss M. L., Hoover E. A. Feline immunodeficiency virus: a neurotropic lentivirus. J Acquir Immune Defic Syndr. 1990;3(7):658–668. [PubMed] [Google Scholar]
  6. Egberink H. F., Ederveen J., Montelaro R. C., Pedersen N. C., Horzinek M. C., Koolen M. J. Intracellular proteins of feline immunodeficiency virus and their antigenic relationship with equine infectious anaemia virus proteins. J Gen Virol. 1990 Mar;71(Pt 3):739–743. doi: 10.1099/0022-1317-71-3-739. [DOI] [PubMed] [Google Scholar]
  7. Elbein A. D., Dorling P. R., Vosbeck K., Horisberger M. Swainsonine prevents the processing of the oligosaccharide chains of influenza virus hemagglutinin. J Biol Chem. 1982 Feb 25;257(4):1573–1576. [PubMed] [Google Scholar]
  8. Elbein A. D., Legler G., Tlusty A., McDowell W., Schwarz R. The effect of deoxymannojirimycin on the processing of the influenza viral glycoproteins. Arch Biochem Biophys. 1984 Dec;235(2):579–588. doi: 10.1016/0003-9861(84)90232-7. [DOI] [PubMed] [Google Scholar]
  9. Elbein A. D., Solf R., Dorling P. R., Vosbeck K. Swainsonine: an inhibitor of glycoprotein processing. Proc Natl Acad Sci U S A. 1981 Dec;78(12):7393–7397. doi: 10.1073/pnas.78.12.7393. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Fuhrmann U., Bause E., Ploegh H. Inhibitors of oligosaccharide processing. Biochim Biophys Acta. 1985 Jun 24;825(2):95–110. doi: 10.1016/0167-4781(85)90095-8. [DOI] [PubMed] [Google Scholar]
  11. Gardner M. B., Luciw P. A. Animal models of AIDS. FASEB J. 1989 Dec;3(14):2593–2606. doi: 10.1096/fasebj.3.14.2556312. [DOI] [PubMed] [Google Scholar]
  12. Gruters R. A., Neefjes J. J., Tersmette M., de Goede R. E., Tulp A., Huisman H. G., Miedema F., Ploegh H. L. Interference with HIV-induced syncytium formation and viral infectivity by inhibitors of trimming glucosidase. Nature. 1987 Nov 5;330(6143):74–77. doi: 10.1038/330074a0. [DOI] [PubMed] [Google Scholar]
  13. Hunsmann G., Moennig V., Pister L., Seifert E., Schäfer W. Properties of mouse leukemia viruses. VIII. The major viral glycoprotein of Friend leukemia virus. Seroimmunological, interfering and hemagglutinating capacities. Virology. 1974 Dec;62(2):307–318. doi: 10.1016/0042-6822(74)90394-8. [DOI] [PubMed] [Google Scholar]
  14. Hunter E., Hill E., Hardwick M., Bhown A., Schwartz D. E., Tizard R. Complete sequence of the Rous sarcoma virus env gene: identification of structural and functional regions of its product. J Virol. 1983 Jun;46(3):920–936. doi: 10.1128/jvi.46.3.920-936.1983. [DOI] [PMC free article] [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. Langlade-Demoyen P., Michel F., Hoffenbach A., Vilmer E., Dadaglio G., Garicia-Pons F., Mayaud C., Autran B., Wain-Hobson S., Plata F. Immune recognition of AIDS virus antigens by human and murine cytotoxic T lymphocytes. J Immunol. 1988 Sep 15;141(6):1949–1957. [PubMed] [Google Scholar]
  17. Lasky L. A., Groopman J. E., Fennie C. W., Benz P. M., Capon D. J., Dowbenko D. J., Nakamura G. R., Nunes W. M., Renz M. E., Berman P. W. Neutralization of the AIDS retrovirus by antibodies to a recombinant envelope glycoprotein. Science. 1986 Jul 11;233(4760):209–212. doi: 10.1126/science.3014647. [DOI] [PubMed] [Google Scholar]
  18. Lenz J., Crowther R., Straceski A., Haseltine W. Nucleotide sequence of the Akv env gene. J Virol. 1982 May;42(2):519–529. doi: 10.1128/jvi.42.2.519-529.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. O'Connor T. P., Jr, Tanguay S., Steinman R., Smith R., Barr M. C., Yamamoto J. K., Pedersen N. C., Andersen P. R., Tonelli Q. J. Development and evaluation of immunoassay for detection of antibodies to the feline T-lymphotropic lentivirus (feline immunodeficiency virus). J Clin Microbiol. 1989 Mar;27(3):474–479. doi: 10.1128/jcm.27.3.474-479.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Oda K., Ikehara Y. Monensin inhibits the conversion of proalbumin to serum albumin in cultured rat hepatocytes. Biochem Biophys Res Commun. 1982 Mar 30;105(2):766–772. doi: 10.1016/0006-291x(82)91500-5. [DOI] [PubMed] [Google Scholar]
  21. Olmsted R. A., Barnes A. K., Yamamoto J. K., Hirsch V. M., Purcell R. H., Johnson P. R. Molecular cloning of feline immunodeficiency virus. Proc Natl Acad Sci U S A. 1989 Apr;86(7):2448–2452. doi: 10.1073/pnas.86.7.2448. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Olmsted R. A., Hirsch V. M., Purcell R. H., Johnson P. R. Nucleotide sequence analysis of feline immunodeficiency virus: genome organization and relationship to other lentiviruses. Proc Natl Acad Sci U S A. 1989 Oct;86(20):8088–8092. doi: 10.1073/pnas.86.20.8088. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Pal R., Hoke G. M., Sarngadharan M. G. Role of oligosaccharides in the processing and maturation of envelope glycoproteins of human immunodeficiency virus type 1. Proc Natl Acad Sci U S A. 1989 May;86(9):3384–3388. doi: 10.1073/pnas.86.9.3384. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Pan Y. T., Hori H., Saul R., Sanford B. A., Molyneux R. J., Elbein A. D. Castanospermine inhibits the processing of the oligosaccharide portion of the influenza viral hemagglutinin. Biochemistry. 1983 Aug 2;22(16):3975–3984. doi: 10.1021/bi00285a038. [DOI] [PubMed] [Google Scholar]
  25. Pedersen N. C., Ho E. W., Brown M. L., Yamamoto J. K. Isolation of a T-lymphotropic virus from domestic cats with an immunodeficiency-like syndrome. Science. 1987 Feb 13;235(4790):790–793. doi: 10.1126/science.3643650. [DOI] [PubMed] [Google Scholar]
  26. Pedersen N. C., Torten M., Rideout B., Sparger E., Tonachini T., Luciw P. A., Ackley C., Levy N., Yamamoto J. Feline leukemia virus infection as a potentiating cofactor for the primary and secondary stages of experimentally induced feline immunodeficiency virus infection. J Virol. 1990 Feb;64(2):598–606. doi: 10.1128/jvi.64.2.598-606.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Pedersen N. C., Yamamoto J. K., Ishida T., Hansen H. Feline immunodeficiency virus infection. Vet Immunol Immunopathol. 1989 May;21(1):111–129. doi: 10.1016/0165-2427(89)90134-7. [DOI] [PubMed] [Google Scholar]
  28. Pinter A., Honnen W. J. O-linked glycosylation of retroviral envelope gene products. J Virol. 1988 Mar;62(3):1016–1021. doi: 10.1128/jvi.62.3.1016-1021.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Pinter A., Honnen W. J. Topography of murine leukemia virus envelope proteins: characterization of transmembrane components. J Virol. 1983 Jun;46(3):1056–1060. doi: 10.1128/jvi.46.3.1056-1060.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Saul R., Chambers J. P., Molyneux R. J., Elbein A. D. Castanospermine, a tetrahydroxylated alkaloid that inhibits beta-glucosidase and beta-glucocerebrosidase. Arch Biochem Biophys. 1983 Mar;221(2):593–597. doi: 10.1016/0003-9861(83)90181-9. [DOI] [PubMed] [Google Scholar]
  31. Saunier B., Kilker R. D., Jr, Tkacz J. S., Quaroni A., Herscovics A. Inhibition of N-linked complex oligosaccharide formation by 1-deoxynojirimycin, an inhibitor of processing glucosidases. J Biol Chem. 1982 Dec 10;257(23):14155–14161. [PubMed] [Google Scholar]
  32. Schultz A. M., Oroszlan S. Tunicamycin inhibits glycosylation of precursor polyprotein encoded by env gene of Rauscher murine leukemia virus. Biochem Biophys Res Commun. 1979 Feb 28;86(4):1206–1213. doi: 10.1016/0006-291x(79)90245-6. [DOI] [PubMed] [Google Scholar]
  33. Srinivas R. V., Melsen L. R., Compans R. W. Effects of monensin on morphogenesis and infectivity of Friend murine leukemia virus. J Virol. 1982 Jun;42(3):1067–1075. doi: 10.1128/jvi.42.3.1067-1075.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Steinman R., Dombrowski J., O'Connor T., Montelaro R. C., Tonelli Q., Lawrence K., Seymour C., Goodness J., Pedersen N. C., Andersen P. R. Biochemical and immunological characterization of the major structural proteins of feline immunodeficiency virus. J Gen Virol. 1990 Mar;71(Pt 3):701–706. doi: 10.1099/0022-1317-71-3-701. [DOI] [PubMed] [Google Scholar]
  35. Stephens E. B., Compans R. W. Assembly of animal viruses at cellular membranes. Annu Rev Microbiol. 1988;42:489–516. doi: 10.1146/annurev.mi.42.100188.002421. [DOI] [PubMed] [Google Scholar]
  36. Stephens E. B., Compans R. W. Nonpolarized expression of a secreted murine leukemia virus glycoprotein in polarized epithelial cells. Cell. 1986 Dec 26;47(6):1053–1059. doi: 10.1016/0092-8674(86)90820-2. [DOI] [PubMed] [Google Scholar]
  37. Stohrer R., Hunter E. Inhibition of Rous sarcoma virus replication by 2-deoxyglucose and tunicamycin: identification of an unglycosylated env gene product. J Virol. 1979 Nov;32(2):412–419. doi: 10.1128/jvi.32.2.412-419.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. Talbott R. L., Sparger E. E., Lovelace K. M., Fitch W. M., Pedersen N. C., Luciw P. A., Elder J. H. Nucleotide sequence and genomic organization of feline immunodeficiency virus. Proc Natl Acad Sci U S A. 1989 Aug;86(15):5743–5747. doi: 10.1073/pnas.86.15.5743. [DOI] [PMC free article] [PubMed] [Google Scholar]
  39. Tartakoff A. M. Perturbation of vesicular traffic with the carboxylic ionophore monensin. Cell. 1983 Apr;32(4):1026–1028. doi: 10.1016/0092-8674(83)90286-6. [DOI] [PubMed] [Google Scholar]
  40. Tartakoff A., Vassalli P. Plasma cell immunoglobulin M molecules. Their biosynthesis, assembly, and intracellular transport. J Cell Biol. 1979 Nov;83(2 Pt 1):284–299. doi: 10.1083/jcb.83.2.284. [DOI] [PMC free article] [PubMed] [Google Scholar]
  41. Tulsiani D. R., Harris T. M., Touster O. Swainsonine inhibits the biosynthesis of complex glycoproteins by inhibition of Golgi mannosidase II. J Biol Chem. 1982 Jul 25;257(14):7936–7939. [PubMed] [Google Scholar]
  42. Veronese F. D., DeVico A. L., Copeland T. D., Oroszlan S., Gallo R. C., Sarngadharan M. G. Characterization of gp41 as the transmembrane protein coded by the HTLV-III/LAV envelope gene. Science. 1985 Sep 27;229(4720):1402–1405. doi: 10.1126/science.2994223. [DOI] [PubMed] [Google Scholar]
  43. Walker B. D., Kowalski M., Goh W. C., Kozarsky K., Krieger M., Rosen C., Rohrschneider L., Haseltine W. A., Sodroski J. Inhibition of human immunodeficiency virus syncytium formation and virus replication by castanospermine. Proc Natl Acad Sci U S A. 1987 Nov;84(22):8120–8124. doi: 10.1073/pnas.84.22.8120. [DOI] [PMC free article] [PubMed] [Google Scholar]
  44. Willey R. L., Bonifacino J. S., Potts B. J., Martin M. A., Klausner R. D. Biosynthesis, cleavage, and degradation of the human immunodeficiency virus 1 envelope glycoprotein gp160. Proc Natl Acad Sci U S A. 1988 Dec;85(24):9580–9584. doi: 10.1073/pnas.85.24.9580. [DOI] [PMC free article] [PubMed] [Google Scholar]
  45. Witte O. N., Wirth D. F. Structure of the murine leukemia virus envelope glycoprotein precursor. J Virol. 1979 Feb;29(2):735–743. doi: 10.1128/jvi.29.2.735-743.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  46. Yamamoto J. K., Sparger E., Ho E. W., Andersen P. R., O'Connor T. P., Mandell C. P., Lowenstine L., Munn R., Pedersen N. C. Pathogenesis of experimentally induced feline immunodeficiency virus infection in cats. Am J Vet Res. 1988 Aug;49(8):1246–1258. [PubMed] [Google Scholar]

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

RESOURCES