The role of Haemagglutinin-Neuraminidase Glycoprotein Cell Surface Expression in the Survival of Sendai Virus-infected BHK-21 Cells

The matrix (M) protein is viewed as the regulator of paramyxovirus particle assembly and budding. Accordingly it was observed to be mutated, and/or decreased in amount, in cases where virus particle production was significantly reduced. Here, a non-productive [nondefective and defective interfering (DI)] Sendai virus infection of COS cells is presented where virus particle production is abolished in the presence of a normal amount of intracellular M protein. In this infection the haemagglutinin-neuraminidase envelope glycoprotein is shown to be dispensable for virion production, and the fusion (F0) envelope glycoprotein behaves as in a productive infection. The M protein is shown to accumulate in perinuclear patches within the cytoplasm. In contrast, localization in the plasma membrane is observed in productive infections. However in both productive and non-productive infections a significant fraction of M protein is found in association with cellular membranes. The M protein-membrane association is shown to take place in the absence of any other viral component, and the M protein-membrane complex exhibits properties similar to those observed for the integral membrane protein F 0. However these properties are distinct from those of the phosphoprotein, which is thought to associate with membranes in a non-specific manner. Concomitant with the cytoplasmic accumulation of M protein and the reduction of virus particle production in this non-productive infection, DI nucleocapsids are shown not to associate with cellular membrane fractions. This is a property which coincides with their poor envelopment in virus particles. Taken together, these data indicate the need for M protein to be recruited at the perinuclear membranes by the nucleocapsids to participate in viral assembly and budding. This view is consistent with a process of viral assembly taking place on internal cytoplasmic membranes rather than at the plasma membrane.

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“…1 a). This is presumably due to more rapid degradation, shown previously in BHK cells (Tuffereau et al, 1985b;Stricker & Roux, 1991).…”

Section: In Cos Cells Sendal Virus Budding Is Restricted In the Presementioning

confidence: 86%

The Sendai Virus Matrix Protein Appears to be Recruited in the Cytoplasm by the Viral Nucleocapsid to function in Viral Assembly and Budding

Stricker

Mottet

Roux

1994

Journal of General Virology

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“…efficiently synthesized on cellular membranes but is degraded during transport to the cell surface (Tuffereau et al, 1985). Mutant virus grown at restrictive temperature exhibits an altered cell tropism -ts271 no longer infects conventional SeV host cells such as HeLa, MDBK and MDCK cells ; however, it still infects human hepatoma HepG2 cells.…”

mentioning

confidence: 99%

Sendai virus-like particles devoid of haemagglutinin-neuraminidase protein infect cells via the human asialoglycoprotein receptor.

Leyrer

Bitzer

Lauer

et al. 1998

Journal of General Virology

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Virus-like particles with genetically defined envelope proteins were generated from cDNA in order to examine the requirement of Sendai virus haemagglutinin-neuraminidase (HN) protein for particle formation, and the role of fusion protein (F) in receptor binding and membrane fusion. Characterization of particles devoid of HN protein showed that particle formation was unimpaired by the absence of HN protein, indicating that HN protein is dispensable for virus assembly and budding. Infection studies further demonstrated that virus adsorption and penetration can be mediated solely by the F protein when the human asialoglycoprotein receptor is present at the surface of host cells.

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“…This surprising result also highlighted the dispensability of HN for viral particle production. This was known for some time due to studies of a thermosensitive mutant (ts271) (13,14) in which HN was degraded at nonpermissive temperatures before reaching the cell membrane (15,16). The finding was then confirmed with the replacement of the SYWST motif by AFYKD, a motif present at the same position of the measles virus (MeV) H protein (7).…”

mentioning

confidence: 48%

Minimal Features of Efficient Incorporation of the Hemagglutinin-Neuraminidase Protein into Sendai Virus Particles

Essaidi-Laziosi

Shevtsova

Roux

2014

J Virol

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Two transmembrane glycoproteins form spikes on the surface of Sendai virus, a member of the Respirovirus genus of the Paramyxovirinae subfamily of the Paramyxoviridae family: the hemagglutinin-neuraminidase (HN) and the fusion (F) proteins. HN, in contrast to F, is dispensable for viral particle production, as normal amounts of particles can be produced with highly reduced levels of HN. This HN reduction can result from mutation of an SYWST motif in its cytoplasmic tail to AFYKD. HN AFYKD accumulates at the infected cell surface but does not get incorporated into particles. In this work, we derived experimental tools to rescue HN AFYKD incorporation. We found that coexpression of a truncated HN harboring the wild-type cytoplasmic tail, the transmembrane domain, and at most 80 amino acids of the ectodomain was sufficient to complement defective HN AFYKD incorporation into particles. This relied on formation of disulfide-bound heterodimers carried out by the two cysteines present in the HN 80-amino-acid (aa) ectodomain. Finally, the replacement of the measles virus H cytoplasmic and transmembrane domains with the corresponding HN domains promoted measles virus H incorporation in Sendai virus particles. Paramyxoviruses are enveloped viruses containing two integral envelope glycoproteins, the hemagglutinin-neuraminidase protein (HN), which is responsible for cell receptor binding/cleavage, and the fusion protein (F), which is responsible for fusion of the viral envelope with the cellular membrane. The inner side of the viral envelope is carpeted by a layer of the matrix M protein that bridges the envelope to the nucleocapsid, the inner core of the particle. The nucleocapsid is composed of a single-stranded RNA of negative polarity, tightly wrapped by nucleocapsid proteins (N) in a structure of helicoidal symmetry, and is associated with the viral RNA-dependent RNA polymerase made of the two proteins P and L (for recent reviews about Paramyxoviridae, see reference 1). In Sendai virus (SeV), a member of the Paramyxovirinae subfamily, genus Respirovirus, HN is a type II glycoprotein of 576 amino acids (aa) having at its N terminus a cytoplasmic tail of 35 aa followed by a transmembrane domain of 25 aa (2). HN has two functions. On one hand, it binds to the sialic acid-containing sugars on cellular glycoproteins, allowing the specific attachment of the virus particle on the cell surface. This in turn triggers the fusion of the viral envelope with the cell membrane effectuated by the F protein. On the other hand, HN contains a neuraminidase (NA) activity, which, by analogy with the influenza virus NA, is likely to allow detachment of the viral particles at the end of the life cycle. HN protrudes from the viral envelope (and is expressed at the cell surface) as a homotetramer composed of two homodimers in which the monomers are covalently linked by disulfide bonds (1, 3, 4). SeV HN protein shares these structural features with the other members of the family. The C-terminal portion of HN forms a globular head that is the ...

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The role of Haemagglutinin-Neuraminidase Glycoprotein Cell Surface Expression in the Survival of Sendai Virus-infected BHK-21 Cells

Cited by 17 publications

References 17 publications

The Sendai Virus Matrix Protein Appears to be Recruited in the Cytoplasm by the Viral Nucleocapsid to function in Viral Assembly and Budding

The Sendai Virus Matrix Protein Appears to be Recruited in the Cytoplasm by the Viral Nucleocapsid to function in Viral Assembly and Budding

Sendai virus-like particles devoid of haemagglutinin-neuraminidase protein infect cells via the human asialoglycoprotein receptor.

Minimal Features of Efficient Incorporation of the Hemagglutinin-Neuraminidase Protein into Sendai Virus Particles

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