1887

Abstract

The role of the glycans of the human immunodeficiency virus type 1 transmembrane glycoprotein (gp41) in the intracellular events of Env precursor (gp160) biosynthesis has been examined by the use of a mutant gp160 in which the cluster of conserved glycosylation sites within the gp41 domain (Asn-621, -630 and -642) has been mutated. Expression of the wild-type and mutant forms of gp160 in BHK-21 cells using recombinant vaccinia viruses has shown that the kinetics of the events occurring in the endoplasmic reticulum (ER) were normal: both Env proteins had similar kinetics of disulphide bond formation, as determined by the acquisition of CD4-binding capability, and both had similar kinetics of oligomer formation. However, in contrast to the parental molecule, mutated gp160 displayed relatively slow transport from the to the medial Golgi where it was retained in the oligomeric state. Transport to the Golgi was impaired, as determined by the sensitivity of gp160 to glycosidases. Cleavage of mutated gp160 at the gp120/gp41 junction was substantially reduced but this was apparently not due to the involvement of the gp41 glycosylation in the cleavage reaction by furin inasmuch as, in the baculovirus system, mutated gp160 could be cleaved when recombinant furin was co-expressed. The reduced cleavage in mammalian cells may thus reflect the impaired routing of mutated Env to the compartment where cleavage occurs. The glycan component of gp41 is, therefore, important for the efficient intracellular transport and processing of gp160. gp160 lacking gp41 carbohydrates is an additional example, among few others, of a protein lacking glycans that is arrested in the Golgi rather than the ER following its biosynthesis.

Loading

Article metrics loading...

/content/journal/jgv/10.1099/0022-1317-76-6-1509
1995-06-01
2024-03-29
Loading full text...

Full text loading...

/deliver/fulltext/jgv/76/6/JV0760061509.html?itemId=/content/journal/jgv/10.1099/0022-1317-76-6-1509&mimeType=html&fmt=ahah

References

  1. Bolmsted A., Hemming A., Flodby P., Bernston P., Travis B., Lin J. P. C., Ledbetter J., Tsu T., Wigzell H., Hu S. L., Olofsson S. 1991; Effects of mutations in glycosylation sites and disulphide bonds on processing, CD4-binding and fusion activity of human immunodeficiency virus envelope glycoproteins. Journal of General Virology 72:1269–1277
    [Google Scholar]
  2. Dash B., McIntosh A., Barrett W., Daniels R. 1994; Deletion of a single N-linked glycosylation site from the transmembrane envelope protein of HIV-1 stops cleavage and transport of gp160 preventing env-mediated fusion. Journal of General Virology 75:1389–1397
    [Google Scholar]
  3. Dedera D., Gu R., Ratner L. 1992; Role of aspargine-linked glycosylation in human immunodeficiency virus type 1 transmembrane envelope function. Virology 187:377–382
    [Google Scholar]
  4. Desrosiers R. C., Tiollais P., Sonigo P. 1987; Sequence of simian immunodeficiency virus from macaque and its relationship to other human and simian immunodeficiency retroviruses. Nature 328:543–547
    [Google Scholar]
  5. Doms R., Lamb R., Rose J. K., Helenius A. 1993; Folding and assembly of viral membrane proteins. Virology 193:545–562
    [Google Scholar]
  6. Earl P. L., Moss B., Doms R. W. 1991; Folding, interaction with GRP78-BiP, assembly, and transport of the human immunodeficiency virus type 1 envelope protein. Journal of Virology 65:2047–2055
    [Google Scholar]
  7. Fenouillet E., Jones I., Powell B., Schmitt D., Kieny M. P., Gluckman J. C. 1993; Functional role of the glycan cluster of HIV-1 transmembrane glycoprotein gp41. Journal of Virology 67:150–160
    [Google Scholar]
  8. Fenouillet E., Gluckman J. C., Jones I. M. 1994; Functions of HIV envelope glycans. Trends in Biochemical Sciences 19:65–70
    [Google Scholar]
  9. Gallaher W. R., Ball J. M., Garry R. F., Griffin M. C., Montelaro R. C. 1989; A general model for the transmembrane proteins of HIV and other retroviruses. AIDS Research and Human Retroviruses 5:431–437
    [Google Scholar]
  10. Guan J. L., Rose J. K. 1984; Conversion of secretory protein into a transmembrane protein results in its transport to the Golgi complex but not to the cell surface. Cell 37:779–787
    [Google Scholar]
  11. Guo H. G., Veronese F., Tschachler E., Pal R., Kalyanaraman V. S., Gallo R. C., Reitz M. 1990; Characterization of an HIV-1 point mutant blocked in envelope glycoprotein cleavage. Virology 174:217–224
    [Google Scholar]
  12. Haffar O. K., Nakamura G. R., Berman P. W. 1990; The carboxy terminus of human immunodeficiency virus type 1 gp160 limits its proteolytic processing and transport in transfected cell lines. Journal of Virology 64:3100–3103
    [Google Scholar]
  13. Hammond C., Braakman I., Helenius A. 1994; Role of N-linked oligosaccharide recognition, glucose trimming and calnexin in glycoprotein folding and quality control. Proceedings of the National Academy of Sciences, USA 91:913–917
    [Google Scholar]
  14. Hurtley S. M. 1992; Golgi localization signals. Trends in Biochemical Sciences 17:2–3
    [Google Scholar]
  15. Hurtley S. M., Helenejs A. 1989; Protein oligomerization in the endoplasmic reticulum. Annual Review of Cell Biology 5:277–307
    [Google Scholar]
  16. Jowett J. B., Jones I. M. 1993; Glycosylation of the external domain of SIV gp41 is influenced by the cytoplasmic tail. AIDS Research and Human Retroviruses 9:197–198
    [Google Scholar]
  17. Kieny M. P., Lathe R., Riviere D., Dott Y., Schmitt K., Girard M., Montagnier L., Lecocq J. P. 1988; Improved antigenicity of the HIV env protein by cleavage site removal. Protein Engineering 2:219–225
    [Google Scholar]
  18. Kitts P., Ayres M., Possee R. 1991; Linearisation of baculovirus DNA improves the selection of recombinant transfer vectors. Nucleic Acids Research 18:5667–5672
    [Google Scholar]
  19. Kornfeld R., Kornfeld S. 1985; Assembly of asparagine-linked oligosaccharides. Annual Review of Biochemistry 5:631–664
    [Google Scholar]
  20. Livingstone C. L., Jones I. M. 1989; Baculovirus expression vectors with single strand capability. Nucleic Acids Research 17:2366
    [Google Scholar]
  21. Machamer C., Florkiewicz R. Z., Rose J. 1985; A single N-linked oligosaccharide at either of the two normal sites is sufficient for transport of vesicular stomatitis virus G protein to the cell surface. Molecular and Cellular Biology 5:3074–3085
    [Google Scholar]
  22. Moore J. P., Jameson B. A., Weiss R. A., Sattentau Q. 1993; The HIV–cell fusion reaction. In Viral Fusion Mechanisms pp 230–289 Edited by Bentz J. Boca Raton: CRC Press;
    [Google Scholar]
  23. Morikawa Y., Barsov E., Jones I. 1993; Legitimate and illegitimate cleavage of human immunodeficiency virus glycoproteins by furin. Journal of Virology 67:3601–3604
    [Google Scholar]
  24. Pal R., Kalyanaraman V. S., Hoke G. M., Sarngadharan M. G. 1989; Processing and secretion of envelope glycoproteins of HIV-1 in the presence of trimming glucosidase inhibitor deoxynojirimycin. Intervirology 30:27–35
    [Google Scholar]
  25. Rose J. K., Doms R. W. 1988; Regulation of protein export from the endoplasmic reticulum. Annual Review of Cell Biology 4:257–288
    [Google Scholar]
  26. Spies C. P., Compans R. 1993; Alternate pathways of secretion of simian immunodeficiency virus envelope glycoproteins. Journal of Virology 67:6535–6541
    [Google Scholar]
  27. Tarentino A. L., Trimble R. B., Plummer T. H. 1989; Enzymatic approaches for studying the structure, synthesis, and processing of glycoproteins. Methods in Cell Biology 32:111–139
    [Google Scholar]
  28. Willey R. L., Klimkait T., Frucht D. M., Bonifacino J. S., Martin D. M. 1991; Mutations within the human immunodeficiency virus type-1 gp160 envelope glycoprotein alter its intracellular transport and processing. Virology 184:326–329
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/jgv/10.1099/0022-1317-76-6-1509
Loading
/content/journal/jgv/10.1099/0022-1317-76-6-1509
Loading

Data & Media loading...

This is a required field
Please enter a valid email address
Approval was a Success
Invalid data
An Error Occurred
Approval was partially successful, following selected items could not be processed due to error