Addition of high-mannose sugars must precede disulfide bond formation for proper folding of Sendai virus glycoproteins

Vidal, S; Mottet, Geneviève; Kolakofsky, Daniel; Roux, Laurent

doi:10.1128/jvi.63.2.892-900.1989

Cited by 70 publications

(28 citation statements)

References 42 publications

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“…6). These latter observations are in agreement with previous experiments involving the HN protein of Sendal virus, in which the acquisition of immunoreactivity with conformation-dependent antibodies was dependent upon glycosylation and the formation of correct intramolecular disulfide bonds (38). Under nonreducing conditions, the d140 and d144 proteins migrated as discrete species of increased electrophoretic mobility ( Fig.…”

Section: The D140 and D144 Proteins Form Authentic Intramolecular Dissupporting

confidence: 92%

Intracellular processing and transport of NH2-terminally truncated forms of a hemagglutinin-neuraminidase type II glycoprotein.

Spriggs

Collins

1990

The Journal of Cell Biology

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Abstract. Six amino-terminal deletion mutants of the NH2-terminally anchored (type II orientation) hemagglutinin-neuraminidase (HN) protein of parainfluenza virus type 3 were expressed in tissue culture by recombinant SV-40 viruses. The mutations consisted of progressive deletions of the cytoplasmic domain and, in some cases, of the hydrophobic signal/anchor. Three activities were dissociated for the signal/anchor: membrane insertion, translocation, and anchoring/transport. HN protein lacking the entire cytoplasmic tail was inserted efficiently into the membrane of the endoplasmic reticulum but was translocated inefficiently into the lumen. However, the small amounts that were successfully translocated appeared to be processed subsequently in a manner indistinguishable from that of parental HN. Thus, the cytoplasmic domain was not required for maturation of this type II glycoprotein. Progressive deletions into the membrane anchor restored efficient translocation, indicating that the NH2-terminal 44 amino acids were fully dispensable for membrane insertion and translocation and that a 10-amino acid hydrophobic signal sequence was sufficient for both activities. These latter HN molecules appeared to be folded authentically as assayed by hemagglutination activity, reactivity with a conformation-specific antiserum, correct formation of intramolecular disulfide bonds, and homooligomerization. However, most (85-90%) of these molecules accumulated in the ER. This showed that folding and oligomerization into a biologically active form, which presumably represents a virion spike, occurs essentially to completion within that compartment but is not sufficient for efficient transport through the exocytotic pathway. Protein transport also appeared to depend on the structure of the membrane anchor. These latter mutants were not stably integrated in the membrane, and the small proportion (10-15%) that was processed through the exocytotic pathway was secreted. The maturation steps and some of the effects of mutations described here for a type II glycoprotein resemble previous observations for prototypic type I glycoproteins and are indicative of close similarities in these processes for proteins of both membrane orientations.T H~ majority of integral membrane glycoproteins span the membrane once and are oriented with either their carboxy-terminal (type I) or amino-terminal (type II) amino acids exposed to the cytoplasm. Type I glycoproteins typically have a cleavable signal sequence at the amino terminus, a carboxy-proximal hydrophobic membrane anchor and a carboxy-terminal hydrophilic cytoplasmic tail. Type II glycoproteins are characterized by an amino-terminal cytoplasmic tail, followed by a 20-30-amino acid hydrophobic transmembrane domain, with the remainder of the protein being oriented externally, comprising the ectodomain.The intracellular processing and transport of membrane glycoproteins has been the object of extensive study. EluciDr. Spriggs's current address is lmmunex Corp., 51 University Street, Seattle WA 98101....

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Section: The D140 and D144 Proteins Form Authentic Intramolecular Dissupporting

confidence: 92%

Intracellular processing and transport of NH2-terminally truncated forms of a hemagglutinin-neuraminidase type II glycoprotein.

Spriggs

Collins

1990

The Journal of Cell Biology

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“…We have previously shown that GSH inhibits replication of parainfluenza-1, herpes simplex-1, and HIV-1 viruses by a direct effect on the envelope glycoproteins (17,31,32). One of the characteristics shared by these proteins is that their assembly into oligomers depends on the formation of disulfide bonds, which are strongly affected by reducing agents such as GSH (66,67). The influenza A virus glycoprotein HA is organized as a homotrimer, and each monomer consists of two disulfide-linked subunits, HA1 and HA2 (54).…”

Section: Discussionmentioning

confidence: 99%

Influenza A virus replication is dependent on an antioxidant pathway that involves GSH and Bcl‐2

Nencioni

Iuvara²,

Aquilano³

et al. 2003

FASEB j.

128

123

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Growing evidence indicates that viral replication is regulated by the redox state of the host cell. We demonstrate that cells of different origins display differential permissivity for influenza A virus replication, depending on their intracellular redox power as reflected by Bcl-2 expression and glutathione (GSH) content. Bcl-2 expressing cells were found to have higher intracellular levels of GSH and to produce lower amounts of virus than Bcl-2 negative cells. Two different steps in the virus life-cycle were involved in Bcl-2/GSH mediated viral inhibition: 1) expression of late viral proteins (in particular hemagglutinin and matrix); and 2) nuclear-cytoplasmic translocation of viral ribonucleoproteins (vRNPs). Buthionine-sulfoximine-induced inhibition of GSH synthesis in Bcl-2 expressing cells caused an increase in the expression of late viral proteins but did not restore vRNP export to the cytoplasm. Collectively, our findings show that both Bcl-2 expression and GSH content contribute to the host cell's ability to down-regulate influenza virus replication, although their effects are exerted at different stages of the viral life-cycle. In certain cell populations, this form of down-regulation might conceivably favor the establishment of persistent viral infection.

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“…This was unexpected because most proteins are glycosylated co-translationally during entry into the ER (2,5,25). Since the folding of some proteins and their transportation from the ER can be greatly affected by glycosylation (24,31), we considered the possibility that the observations made with VP7 might be connected with the mechanism by which this protein is retained in the ER. In this work, we examine the kinetics of VP7 glycosylation in more detail and investigate the reason for its delay.…”

mentioning

confidence: 99%

Sequences in rotavirus glycoprotein VP7 that mediate delayed translocation and retention of the protein in the endoplasmic reticulum.

Stirzaker¹,

Poncet²,

Both³

1990

The Journal of Cell Biology

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Abstract. Glycosylation and translocation of the simian rotavirus protein VP7, a resident ER protein, does not occur co-translationally in vivo. In pulse-chase experiments in COS cells, nonglycosylated VP7 was still detectable after a 25-min chase period, although the single glycosylation site was only 18 residues beyond the signal peptide cleavage site. After labeling, glycosylated and nonglycosylated VP7 was recovered in microsomes but the latter was sensitive to trypsin (i.e., the nascent protein became membrane associated) but most of it entered the ER posttranslationally because of a rate-limiting step early in translocation. In contrast with the simian protein, bovine VP7 was glycosylated and translocated rapidly. Thus, delayed translocation per se was not required for retention of VP7 in the ER. By constructing hybrid proteins, it was further shown that the signal peptide together with residues 64-111 of the simian protein caused delayed translocation. The same sequences were also necessary and sufficient for retention of simian VP7 in the ER. The data are consistent with the idea that certain proteins are inserted into the ER membrane in a loop configuration.OTAVIRUSES, as members of the family Reoviridae, have a segmented double-stranded (ds) ~ RNA genome and replicate in the cytoplasm of infected cells. Electron microscopy has shown that viral morphogenesis begins in the cytoplasm in viroplasmic inclusions where the central core and single-shelled particles are assembled (for review see references 4 and 13). The latter enter the ER during a budding process that is mediated by the nonstructural, resident ER glycoprotein NS28 (3, 4). The outer capsid layer of mature virus particles consists of VP4 and the glycoprotein VP7 (14), but it is not clear how these are acquired. An inspection of VP4 sequences (7,20) suggests that the protein has no signal peptide and presumably enters the ER as part of the budding particle. However, VP7 contains a cleavable signal peptide that directs it to the ER independently of the single-shelled particles (12,29,32). Analysis of the carbohydrate attached to VP7 shows that it is of the high-mannose type (18) indicating that the protein is retained in the ER where virus maturation occurs. This has been confirmed by the expression of cloned VP7 genes in COS cells (23,28). Other studies have also shown that VP7 is retained in the ER as an integral membrane protein (18).The rotavirus VP7 signal peptide (referred to as H2; reference 32) is involved in directing the protein to the ER and retaining it there. This was shown by replacing the H2 signal sequence with the signal peptide from influenza virus

show abstract

Addition of high-mannose sugars must precede disulfide bond formation for proper folding of Sendai virus glycoproteins

Cited by 70 publications

References 42 publications

Intracellular processing and transport of NH2-terminally truncated forms of a hemagglutinin-neuraminidase type II glycoprotein.

Intracellular processing and transport of NH2-terminally truncated forms of a hemagglutinin-neuraminidase type II glycoprotein.

Influenza A virus replication is dependent on an antioxidant pathway that involves GSH and Bcl‐2

Sequences in rotavirus glycoprotein VP7 that mediate delayed translocation and retention of the protein in the endoplasmic reticulum.

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