Synthesis and Assembly of Viral Membrane Proteins*

Lodish, Harvey F.; Wirth, Dyann F.; Porter, Mary

doi:10.1111/j.1749-6632.1980.tb47261.x

Cited by 6 publications

(4 citation statements)

References 48 publications

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“…We do not know whether these virus-like particles utilize cellular glycoproteins in their assembly, but it appears that under our conditions of infection, HSV fails to insert non-glycosylated forms of glycoproteins into membranes. The few nucleocapsids which can be observed may be budding through those patches of membrane which have been modified by the input virus, or they may be using cellular glycoproteins for assembly (23). The virions on the surface could be partially uncoated virions remaining from the infecting virus.…”

Section: Discussionmentioning

confidence: 99%

Herpesvirus glycoprotein synthesis and insertion into plasma membranes

Peake¹,

Nystrom²,

Pizer³

1982

J Virol

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In the presence of the antibiotic tunicamycin (TM), glycosylation of herpes simplex virus glycoproteins is inhibited and non-glycosylated polypeptides analogous to the glycoproteins are synthesized (Pizer et al., J. Virol. 34:142-153, 1980). The synthesis of viral proteins and DNA occurs in TM-treated cells. By electron microscopy, nucleocapsids can be observed both in the nucleus and the cytoplasm of TM-treated cells; a small number of enveloped virions were observed on the cell surface. Analyses of the proteins in partially purified virus readily detects viral glycoproteins in the control cells, but neither glycoproteins nor nonglycosylated polypeptide analogs were observed in the virus prepared from TM-treated cells. By labeling the surface of infected cells with 125I, viral glycoproteins were detected as soon as 90 min after infection even when protein synthesis was inhibited with cycloheximide and glycosylation was blocked with TM. Labeling the proteins synthesized in infected cells with [35S]methionine showed that the surface glycoproteins detected in the cycloheximide- and TM-treated cells were not synthesized de novo after infection, but were placed on the cell surface by the infecting virus. Studies with metabolic inhibitors and a temperature-sensitive mutant blocked early in the infectious cycle showed that glycoproteins gA/gB and gD were synthesized soon after infection, but that the synthesis of gC was delayed. Under conditions of infection, in which gC and its precursor pgC are not produced, we have been able to observe the relationships between the glycosylated polypeptides that correspond to pgA/pgB and the nonglycosylated analog made in the presence of TM.

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Section: Discussionmentioning

confidence: 99%

Herpesvirus glycoprotein synthesis and insertion into plasma membranes

Peake¹,

Nystrom²,

Pizer³

1982

J Virol

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“…The vesicular stomatitis viral glycoprotein, G, like cellsurface glycoproteins, is cotranslationally glycosylated and inserted into the rough endoplasmic reticulum membrane (RER, reviewed by Sabatini et al, 1982;Wickner and Lodish, 1985). From the RER it is transported first to the Golgi complex (Bergmann et al, 1981;Wehland et a/., 1982) and then to the plasma membrane, where assembly and budding of viral particles occurs (reviewed by Lenard and Compans, 1974;Atkinson, 1978;Lodish et al, 1980). In the same cell, different membrane and secreted proteins can take very different times for transport to the cell surface; the rate-limiting and distinctive step appears to be transport from the RER to an early (or medial) Golgi compartment (Fitting and Kabat, 1982;Ledford and Davis, 1983;Williams et al, 1985).…”

mentioning

confidence: 99%

Oligomerization is essential for transport of vesicular stomatitis viral glycoprotein to the cell surface

Kreis¹,

Lodish²

1986

Cell

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409

284

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Using ts045, a temperature sensitive strain of Vesicular stomatitis virus, we show that oligomerization of G protein is a prerequisite for its transport from RER to the Golgi apparatus and for its subsequent maturation. While wild-type G forms an oligomer in the RER, ts045 G synthesized at the nonpermissive temperature does not. When the permissive temperature is reinstated, ts045 G forms an oligomer and moves to the Golgi. The state of oligomerization was determined by chemical cross-linking and by the ability of a microinjected monoclonal antibody specific for the carboxy-terminal five amino acids of the cytoplasmic tail of G to cause patching of G in intracellular membranes. We conclude that formation of an oligomer of G protein, probably a trimer, is necessary for G protein maturation.

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“…Glycoprotein-specific antisera have been used to detect at least one partially glycosylated intermediate for each major glycoprotein species (Spear, 1976;Eisenberg et al, 1979;Eberle and Courtney, 1980b). Based on studies of vesicular stomatitis virus (VSV) and Sindbis virus glycoprotein processing and intracellular transport (Hunt et al, 1978;Katz et al, 1977;Knipe et al, 1977;Lodish et al, 1980;Rothman et al, 1980a, b), it might be assumed that the HSV glycoprotein messenger RNAs are translated on membranebound polysomes and that a mannose-rich oligosaccharide core becomes covalently attached to specific asparagine residues by rough endoplasmic reticulum-associated glycosyl transferases as a cotranslational event. Pizer et al (1980) have provided evidence for N-linked mannose core glycosylation of glycoprotein gD.…”

mentioning

confidence: 99%

Inhibition of glycosylation of herpes simplex virus glycoproteins: Identification of antigenic and immunogenic partially glycosylated glycopeptides on the cell surface membrane

et al. 1983

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The surface membranes of cells infected with herpes simplex virus type 1 (HSV-1), strain KOS, contain three principal glycoproteins, gC (apparent Mr 129k), gB (apparent Mr 120k), and gD (apparent Mr 58k). Infections carried out in the presence of the glycosylation inhibitor 2-deoxy-D-glucose result in the loss of the mature species with the concurrent appearance of lower-molecular-weight polypeptides which are presumably partially glycosylated forms of the fully processed glycoproteins. Specific immunoprecipitation of radiolabeled cytoplasmic extracts of 2-deoxy-D-glucose-inhibited infections identified partially glycosylated proteins designated DG92, DG88, and DG53, which are antigenically related to the corresponding mature forms gB, gC, and gD. Cell surface radioiodination, in combination with specific immunoprecipitation, revealed that DG88 and DG53 were the principal species transported to the cell surface in 2-deoxy-D-glucose-inhibited infections. DG92 was readily detected in the cytoplasm but not on the plasma membrane. Cells infected with the KOS mutant, syn LD70, did not synthesize glycoprotein gC. In glycosylation-inhibited syn LD70 infections, DG88 was not detected in either the cytoplasm or plasma membrane, demonstrating a genetic relationship between DG88 and gC. Polyclonal and monoclonal antibodies directed against the glycoproteins gC, gB, and gD sensitized infected cells to complement-mediated immune cytolysis. Cells infected in the presence of the inhibitor were sensitized to lysis only by antibody specific for gC and gD. The glycosylation-inhibited cells were insensitive to immunolysis by anti-gB monoclonal antibody. These findings confirm that the glycosylation-deficient forms of gC and gD, but not gB reach the cell surface in the presence of inhibitor and that the inhibitor-induced alterations in glycosylation do not cause a complete loss of antigenicity. Inoculation of mice with syngeneic 3T3 cells infected in the presence or absence of inhibitor-induced cytolytic and neutralizing antibody. A major portion of the cytolytic antibody was directed against gC, but anti-gC antibody appeared to play a minor role in virus neutralization. While the serum induced by the control infected cells contained precipitating antibodies for gC, gB, and gD, the serum derived from mice inoculated with inhibitor-treated infected cells had only weak immunoprecipitating activity against gB. Together, these findings have identified partially glycosylated forms of the major HSV glycoproteins and show that complete glycosylation is not required for transport of some of these partially glycosylated polypeptides to the cell surface. Moreover, complete glycosylation of the glycopeptides is not essential for maintenance of antigenicity or immunogenicity, indicating that at least some determinants recognized by antibodies directed against the mature glycoproteins are not affected by 2-deoxy-D-glucose-induced carbohydrate alterations.

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Synthesis and Assembly of Viral Membrane Proteins*

Cited by 6 publications

References 48 publications

Herpesvirus glycoprotein synthesis and insertion into plasma membranes

Herpesvirus glycoprotein synthesis and insertion into plasma membranes

Oligomerization is essential for transport of vesicular stomatitis viral glycoprotein to the cell surface

Inhibition of glycosylation of herpes simplex virus glycoproteins: Identification of antigenic and immunogenic partially glycosylated glycopeptides on the cell surface membrane

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