Skip to main content
Access keys NCBI Homepage MyNCBI Homepage Main Content Main Navigation
J Virol. 1991 Apr; 65(4): 2047–2055.
doi: 10.1128/jvi.65.4.2047-2055.1991
PMCID: PMC240054
PMID: 1900540

Folding, interaction with GRP78-BiP, assembly, and transport of the human immunodeficiency virus type 1 envelope protein.

P L Earl, B Moss, and R W Doms
Author information Copyright and License information PMC Disclaimer

Abstract

A detailed kinetic and quantitative analysis of the early and late biosynthetic events undergone by the human immunodeficiency virus type 1 envelope protein expressed by a recombinant vaccinia virus was performed. Early folding events that occurred in the endoplasmic reticulum included disulfide bond formation (t1/2 approximately 10 min), folding of envelope protein into a form competent to bind CD4 (t1/2 approximately 15 min), and specific and transient association and dissociation with GRP78-BiP (t1/2 approximately 25 min). After initial folding, envelope protein monomers formed noncovalently associated dimers with high efficiency (t1/2 approximately 30 min). Studies with brefeldin A, a compound that inhibits endoplasmic reticulum-to-Golgi transport, suggested that assembly occurred in the endoplasmic reticulum while cleavage of gp160 into gp120/gp41 subunits occurred in a post-endoplasmic reticulum compartment. Transport to the Golgi was monitored by modification of N-linked sugars to forms partially resistant to endoglycosidase H. The kinetics of endoglycosidase H resistance were nearly identical to the kinetics of gp160 cleavage (t1/2 approximately 80 min). Cleavage efficiency was strongly cell type dependent, ranging from 13 to 70%. By contrast, approximately 50% of the gp120 generated by the cleavage event was shed (t1/2 approximately 120 min) regardless of the cell type used. The results are discussed in terms of the overall biosynthetic pathway of the envelope protein and provide a framework with which to assess the effects of mutations on structure and function.

Full text

Full text is available as a scanned copy of the original print version. Get a printable copy (PDF file) of the complete article (2.0M), or click on a page image below to browse page by page. Links to PubMed are also available for Selected References.

 
icon of scanned page 2047
2047
icon of scanned page 2048
2048
icon of scanned page 2049
2049
icon of scanned page 2050
2050
icon of scanned page 2051
2051
icon of scanned page 2052
2052
icon of scanned page 2053
2053
icon of scanned page 2054
2054
icon of scanned page 2055
2055
 

Images in this article

Image

Image
on p.2048

Image

Image
on p.2050

Image

Image
on p.2050

Image

Image
on p.2051

Image

Image
on p.2051

Image

Image
on p.2052

Click on the image to see a larger version.

Selected References

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

  • Allan JS, Coligan JE, Barin F, McLane MF, Sodroski JG, Rosen CA, Haseltine WA, Lee TH, Essex M. Major glycoprotein antigens that induce antibodies in AIDS patients are encoded by HTLV-III. Science. 1985 May 31;228(4703):1091–1094. [PubMed] [Google Scholar]
  • Aroeti B, Henis YI. Effects of fusion temperature on the lateral mobility of Sendai virus glycoproteins in erythrocyte membranes and on cell fusion indicate that glycoprotein mobilization is required for cell fusion. Biochemistry. 1988 Jul 26;27(15):5654–5661. [PubMed] [Google Scholar]
  • Bole DG, Hendershot LM, Kearney JF. Posttranslational association of immunoglobulin heavy chain binding protein with nascent heavy chains in nonsecreting and secreting hybridomas. J Cell Biol. 1986 May;102(5):1558–1566. [PMC free article] [PubMed] [Google Scholar]
  • Bosch ML, Earl PL, Fargnoli K, Picciafuoco S, Giombini F, Wong-Staal F, Franchini G. Identification of the fusion peptide of primate immunodeficiency viruses. Science. 1989 May 12;244(4905):694–697. [PubMed] [Google Scholar]
  • Boulay F, Doms RW, Webster RG, Helenius A. Posttranslational oligomerization and cooperative acid activation of mixed influenza hemagglutinin trimers. J Cell Biol. 1988 Mar;106(3):629–639. [PMC free article] [PubMed] [Google Scholar]
  • Buonocore L, Rose JK. Prevention of HIV-1 glycoprotein transport by soluble CD4 retained in the endoplasmic reticulum. Nature. 1990 Jun 14;345(6276):625–628. [PubMed] [Google Scholar]
  • Ceriotti A, Colman A. Trimer formation determines the rate of influenza virus haemagglutinin transport in the early stages of secretion in Xenopus oocytes. J Cell Biol. 1990 Aug;111(2):409–420. [PMC free article] [PubMed] [Google Scholar]
  • Chakrabarti S, Mizukami T, Franchini G, Moss B. Synthesis, oligomerization, and biological activity of the human immunodeficiency virus type 2 envelope glycoprotein expressed by a recombinant vaccinia virus. Virology. 1990 Sep;178(1):134–142. [PubMed] [Google Scholar]
  • Chege NW, Pfeffer SR. Compartmentation of the Golgi complex: brefeldin-A distinguishes trans-Golgi cisternae from the trans-Golgi network. J Cell Biol. 1990 Sep;111(3):893–899. [PMC free article] [PubMed] [Google Scholar]
  • Copeland CS, Doms RW, Bolzau EM, Webster RG, Helenius A. Assembly of influenza hemagglutinin trimers and its role in intracellular transport. J Cell Biol. 1986 Oct;103(4):1179–1191. [PMC free article] [PubMed] [Google Scholar]
  • Crise B, Ruusala A, Zagouras P, Shaw A, Rose JK. Oligomerization of glycolipid-anchored and soluble forms of the vesicular stomatitis virus glycoprotein. J Virol. 1989 Dec;63(12):5328–5333. [PMC free article] [PubMed] [Google Scholar]
  • Daniels RS, Downie JC, Hay AJ, Knossow M, Skehel JJ, Wang ML, Wiley DC. Fusion mutants of the influenza virus hemagglutinin glycoprotein. Cell. 1985 Feb;40(2):431–439. [PubMed] [Google Scholar]
  • Dewar RL, Vasudevachari MB, Natarajan V, Salzman NP. 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. [PMC free article] [PubMed] [Google Scholar]
  • Doms RW, Earl PL, Chakrabarti S, Moss B. Human immunodeficiency virus types 1 and 2 and simian immunodeficiency virus env proteins possess a functionally conserved assembly domain. J Virol. 1990 Jul;64(7):3537–3540. [PMC free article] [PubMed] [Google Scholar]
  • Doms RW, Gething MJ, Henneberry J, White J, Helenius A. Variant influenza virus hemagglutinin that induces fusion at elevated pH. J Virol. 1986 Feb;57(2):603–613. [PMC free article] [PubMed] [Google Scholar]
  • Doms RW, Keller DS, Helenius A, Balch WE. Role for adenosine triphosphate in regulating the assembly and transport of vesicular stomatitis virus G protein trimers. J Cell Biol. 1987 Nov;105(5):1957–1969. [PMC free article] [PubMed] [Google Scholar]
  • Doms RW, Russ G, Yewdell JW. Brefeldin A redistributes resident and itinerant Golgi proteins to the endoplasmic reticulum. J Cell Biol. 1989 Jul;109(1):61–72. [PMC free article] [PubMed] [Google Scholar]
  • Dorner AJ, Bole DG, Kaufman RJ. The relationship of N-linked glycosylation and heavy chain-binding protein association with the secretion of glycoproteins. J Cell Biol. 1987 Dec;105(6 Pt 1):2665–2674. [PMC free article] [PubMed] [Google Scholar]
  • Earl PL, Doms RW, Moss B. Oligomeric structure of the human immunodeficiency virus type 1 envelope glycoprotein. Proc Natl Acad Sci U S A. 1990 Jan;87(2):648–652. [PMC free article] [PubMed] [Google Scholar]
  • Earl PL, Hügin AW, Moss B. Removal of cryptic poxvirus transcription termination signals from the human immunodeficiency virus type 1 envelope gene enhances expression and immunogenicity of a recombinant vaccinia virus. J Virol. 1990 May;64(5):2448–2451. [PMC free article] [PubMed] [Google Scholar]
  • Earl PL, Koenig S, Moss B. Biological and immunological properties of human immunodeficiency virus type 1 envelope glycoprotein: analysis of proteins with truncations and deletions expressed by recombinant vaccinia viruses. J Virol. 1991 Jan;65(1):31–41. [PMC free article] [PubMed] [Google Scholar]
  • Einfeld D, Hunter E. Oligomeric structure of a prototype retrovirus glycoprotein. Proc Natl Acad Sci U S A. 1988 Nov;85(22):8688–8692. [PMC free article] [PubMed] [Google Scholar]
  • Fennie C, Lasky LA. Model for intracellular folding of the human immunodeficiency virus type 1 gp120. J Virol. 1989 Feb;63(2):639–646. [PMC free article] [PubMed] [Google Scholar]
  • Freed EO, Myers DJ, Risser R. Characterization of the fusion domain of the human immunodeficiency virus type 1 envelope glycoprotein gp41. Proc Natl Acad Sci U S A. 1990 Jun;87(12):4650–4654. [PMC free article] [PubMed] [Google Scholar]
  • Gething MJ, Doms RW, York D, White J. Studies on the mechanism of membrane fusion: site-specific mutagenesis of the hemagglutinin of influenza virus. J Cell Biol. 1986 Jan;102(1):11–23. [PMC free article] [PubMed] [Google Scholar]
  • Geyer H, Holschbach C, Hunsmann G, Schneider J. Carbohydrates of human immunodeficiency virus. Structures of oligosaccharides linked to the envelope glycoprotein 120. J Biol Chem. 1988 Aug 25;263(24):11760–11767. [PubMed] [Google Scholar]
  • Hurtley SM, Helenius A. Protein oligomerization in the endoplasmic reticulum. Annu Rev Cell Biol. 1989;5:277–307. [PubMed] [Google Scholar]
  • Kowalski M, Potz J, Basiripour L, Dorfman T, Goh WC, Terwilliger E, Dayton A, Rosen C, Haseltine W, Sodroski J. Functional regions of the envelope glycoprotein of human immunodeficiency virus type 1. Science. 1987 Sep 11;237(4820):1351–1355. [PubMed] [Google Scholar]
  • Lippincott-Schwartz J, Yuan LC, Bonifacino JS, Klausner RD. Rapid redistribution of Golgi proteins into the ER in cells treated with brefeldin A: evidence for membrane cycling from Golgi to ER. Cell. 1989 Mar 10;56(5):801–813. [PMC free article] [PubMed] [Google Scholar]
  • Machamer CE, Doms RW, Bole DG, Helenius A, Rose JK. Heavy chain binding protein recognizes incompletely disulfide-bonded forms of vesicular stomatitis virus G protein. J Biol Chem. 1990 Apr 25;265(12):6879–6883. [PubMed] [Google Scholar]
  • McCune JM, Rabin LB, Feinberg MB, Lieberman M, Kosek JC, Reyes GR, Weissman IL. Endoproteolytic cleavage of gp160 is required for the activation of human immunodeficiency virus. Cell. 1988 Apr 8;53(1):55–67. [PubMed] [Google Scholar]
  • McKeating JA, Willey RL. Structure and function of the HIV envelope. AIDS. 1989;3 (Suppl 1):S35–S41. [PubMed] [Google Scholar]
  • Mizukami T, Fuerst TR, Berger EA, Moss B. Binding region for human immunodeficiency virus (HIV) and epitopes for HIV-blocking monoclonal antibodies of the CD4 molecule defined by site-directed mutagenesis. Proc Natl Acad Sci U S A. 1988 Dec;85(23):9273–9277. [PMC free article] [PubMed] [Google Scholar]
  • Mizuochi T, Matthews TJ, Kato M, Hamako J, Titani K, Solomon J, Feizi T. Diversity of oligosaccharide structures on the envelope glycoprotein gp 120 of human immunodeficiency virus 1 from the lymphoblastoid cell line H9. Presence of complex-type oligosaccharides with bisecting N-acetylglucosamine residues. J Biol Chem. 1990 May 25;265(15):8519–8524. [PubMed] [Google Scholar]
  • Ng DT, Randall RE, Lamb RA. Intracellular maturation and transport of the SV5 type II glycoprotein hemagglutinin-neuraminidase: specific and transient association with GRP78-BiP in the endoplasmic reticulum and extensive internalization from the cell surface. J Cell Biol. 1989 Dec;109(6 Pt 2):3273–3289. [PMC free article] [PubMed] [Google Scholar]
  • Parks GD, Lamb RA. Defective assembly and intracellular transport of mutant paramyxovirus hemagglutinin-neuraminidase proteins containing altered cytoplasmic domains. J Virol. 1990 Aug;64(8):3605–3616. [PMC free article] [PubMed] [Google Scholar]
  • Parks GD, Lamb RA. Folding and oligomerization properties of a soluble and secreted form of the paramyxovirus hemagglutinin-neuraminidase glycoprotein. Virology. 1990 Oct;178(2):498–508. [PubMed] [Google Scholar]
  • Pinter A, Honnen WJ, Tilley SA, Bona C, Zaghouani H, Gorny MK, Zolla-Pazner S. Oligomeric structure of gp41, the transmembrane protein of human immunodeficiency virus type 1. J Virol. 1989 Jun;63(6):2674–2679. [PMC free article] [PubMed] [Google Scholar]
  • Ratner L, Haseltine W, Patarca R, Livak KJ, Starcich B, Josephs SF, Doran ER, Rafalski JA, Whitehorn EA, Baumeister K, et al. Complete nucleotide sequence of the AIDS virus, HTLV-III. Nature. 1985 Jan 24;313(6000):277–284. [PubMed] [Google Scholar]
  • Rose JK, Doms RW. Regulation of protein export from the endoplasmic reticulum. Annu Rev Cell Biol. 1988;4:257–288. [PubMed] [Google Scholar]
  • Roux L. Selective and transient association of Sendai virus HN glycoprotein with BiP. Virology. 1990 Mar;175(1):161–166. [PubMed] [Google Scholar]
  • Schawaller M, Smith GE, Skehel JJ, Wiley DC. Studies with crosslinking reagents on the oligomeric structure of the env glycoprotein of HIV. Virology. 1989 Sep;172(1):367–369. [PubMed] [Google Scholar]
  • Singh I, Doms RW, Wagner KR, Helenius A. Intracellular transport of soluble and membrane-bound glycoproteins: folding, assembly and secretion of anchor-free influenza hemagglutinin. EMBO J. 1990 Mar;9(3):631–639. [PMC free article] [PubMed] [Google Scholar]
  • Stegmann T, Doms RW, Helenius A. Protein-mediated membrane fusion. Annu Rev Biophys Biophys Chem. 1989;18:187–211. [PubMed] [Google Scholar]
  • Stein BS, Engleman EG. Intracellular processing of the gp160 HIV-1 envelope precursor. Endoproteolytic cleavage occurs in a cis or medial compartment of the Golgi complex. J Biol Chem. 1990 Feb 15;265(5):2640–2649. [PubMed] [Google Scholar]
  • Veronese FD, DeVico AL, Copeland TD, Oroszlan S, Gallo RC, Sarngadharan MG. Characterization of gp41 as the transmembrane protein coded by the HTLV-III/LAV envelope gene. Science. 1985 Sep 27;229(4720):1402–1405. [PubMed] [Google Scholar]
  • Waxham MN, Merz DC, Wolinsky JS. Intracellular maturation of mumps virus hemagglutinin-neuraminidase glycoprotein: conformational changes detected with monoclonal antibodies. J Virol. 1986 Aug;59(2):392–400. [PMC free article] [PubMed] [Google Scholar]
  • Weiss CD, Levy JA, White JM. Oligomeric organization of gp120 on infectious human immunodeficiency virus type 1 particles. J Virol. 1990 Nov;64(11):5674–5677. [PMC free article] [PubMed] [Google Scholar]
  • White JM. Viral and cellular membrane fusion proteins. Annu Rev Physiol. 1990;52:675–697. [PubMed] [Google Scholar]
  • Willey RL, Bonifacino JS, Potts BJ, Martin MA, Klausner RD. 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. [PMC free article] [PubMed] [Google Scholar]
  • Yewdell JW, Yellen A, Bächi T. Monoclonal antibodies localize events in the folding, assembly, and intracellular transport of the influenza virus hemagglutinin glycoprotein. Cell. 1988 Mar 25;52(6):843–852. [PubMed] [Google Scholar]
Articles from Journal of Virology are provided here courtesy of American Society for Microbiology (ASM)
-