Analysis of Respiratory Syncytial Virus F, G, and SH Proteins in Cell Fusion

Heminway, B. R.; Yu, Yang; Tanaka, Yoshikazu; Perrine, Kimberly G.; Gustafson, Elisabet; Bernstein, Jack; Galinski, Mark S.

doi:10.1006/viro.1994.1245

Cited by 110 publications

(85 citation statements)

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“…Syncytium formation was observed in both recWT virus-and M-null virus-infected cell cultures. This was not surprising, since the expression of the F protein alone is capable of inducing extensive cell-cell membrane fusion (5,26). In the absence of M, however, syncytia were limited in size, and healthy uninfected cells in the culture kept dividing throughout the analysis.…”

Section: Discussionmentioning

confidence: 98%

“…G is a highly glycosylated protein that is expressed as a secreted form and a membraneanchored form, with the latter serving as a viral attachment protein (34). F resembles the prototypic paramyxovirus fusion protein but can induce membrane fusion in the absence of G (26,39). F also appears to play a role in viral attachment, and nucleolin was recently identified as a cellular receptor for the F protein (57).…”

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confidence: 99%

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The Human Respiratory Syncytial Virus Matrix Protein Is Required for Maturation of Viral Filaments

Mitra

Baviskar

Duncan-Decocq

et al. 2012

J Virol

110

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An experimental system was developed to generate infectious human respiratory syncytial virus (HRSV) lacking matrix (M) protein expression (M-null virus) from cDNA. The role of the M protein in virus assembly was then examined by infecting HEp-2 and Vero cells with the M-null virus and assessing the impact on infectious virus production and viral protein trafficking. In the absence of M, the production of infectious progeny was strongly impaired. Immunofluorescence (IF) microscopy analysis using antibodies against the nucleoprotein (N), attachment protein (G), and fusion protein (F) failed to detect the characteristic virusinduced cell surface filaments, which are believed to represent infectious virions. In addition, a large proportion of the N protein was detected in viral replication factories termed inclusion bodies (IBs). High-resolution analysis of the surface of M-null virusinfected cells by field emission scanning electron microscopy (SEM) revealed the presence of large areas with densely packed, uniformly short filaments. Although unusually short, these filaments were otherwise similar to those induced by an M-containing control virus, including the presence of the viral G and F proteins. The abundance of the short, stunted filaments in the absence of M indicates that M is not required for the initial stages of filament formation but plays an important role in the maturation or elongation of these structures. In addition, the absence of mature viral filaments and the simultaneous increase in the level of the N protein within IBs suggest that the M protein is involved in the transport of viral ribonucleoprotein (RNP) complexes from cytoplasmic IBs to sites of budding.

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Section: Discussionmentioning

confidence: 98%

mentioning

confidence: 99%

The Human Respiratory Syncytial Virus Matrix Protein Is Required for Maturation of Viral Filaments

Mitra

Baviskar

Duncan-Decocq

et al. 2012

J Virol

110

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“…It has been postulated that the interaction of paramyxovirus attachment protein with the cell receptor triggers a conformational change in the F protein that promotes membrane fusion (28). It has also been reported that the G and SH glycoproteins of HRSV enhance the fusion activity of F proteins when coexpressed in transfected cells (29). However, a deletion mutant of HRSV lacking both the G and SH genes is able to replicate efficiently in certain cell lines and to produce syncytia (30).…”

Section: Discussionmentioning

confidence: 99%

Cleavage of the human respiratory syncytial virus fusion protein at two distinct sites is required for activation of membrane fusion

González-Reyes

Ruiz-Argüello

Garcı́a-Barreno

et al. 2001

Proc. Natl. Acad. Sci. U.S.A.

206

175

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Preparations of purified full-length fusion (F) protein of human respiratory syncytial virus (HRSV) expressed in recombinant vaccinia-F infected cells, or of an anchorless mutant (F TM؊) lacking the C-terminal 50 amino acids secreted from vaccinia-F TM؊-infected cells contain a minor polypeptide that is an intermediate product of proteolytic processing of the F protein precursor F0. N-terminal sequencing of the intermediate demonstrated that it is generated by cleavage at a furin-motif, residues 106 -109 of the F sequence. By contrast, the F1 N terminus derives from cleavage at residue 137 of F0 which is also C-terminal to a furin recognition site at residues 131-136. Site-directed mutagenesis indicates that processing of F0 protein involves independent cleavage at both sites. Both cleavages are required for the F protein to be active in membrane fusion as judged by syncytia formation, and they allow changes in F structure from cone-to lollipop-shaped spikes and the formation of rosettes by anchorless F.

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“…The SH protein of HRSV exists as different forms, SHg (13-15 kDa), SHp (21-30 kDa), non-glycosylated SH0 (7.5 kDa) and a nonglycosylated form SHt (4.8 kDa) which is derived from initiation at a second methionine in the open reading frame (ORF) [4,111]. There is evidence that the SH protein may play a role in virusmediated cell fusion by interacting with the F protein [43,59].…”

Section: Small Hydrophobic Sh Proteinmentioning

confidence: 99%

Bovine respiratory syncytial virus infection

Valarcher¹,

2007

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-Bovine respiratory syncytial virus (BRSV) belongs to the pneumovirus genus within the family Paramyxoviridae and is a major cause of respiratory disease in young calves. BRSV is enveloped and contains a negative sense, single-stranded RNA genome encoding 11 proteins. The virus replicates predominantly in ciliated respiratory epithelial cells but also in type II pneumocytes. It appears to cause little or no cytopathology in ciliated epithelial cell cultures in vitro, suggesting that much of the pathology is due to the host's response to virus infection. RSV infection induces an array of pro-inflammatory chemokines and cytokines that recruit neutrophils, macrophages and lymphocytes to the respiratory tract resulting in respiratory disease. Although the mechanisms responsible for induction of these chemokines and cytokines are unclear, studies on the closely related human (H)RSV suggest that activation of NF-κB via TLR4 and TLR3 signalling pathways is involved. An understanding of the mechanisms by which BRSV is able to establish infection and induce an inflammatory response has been facilitated by advances in reverse genetics, which have enabled manipulation of the virus genome. These studies have demonstrated an important role for the non-structural proteins in anti-interferon activity, a role for a virokinin, released during proteolytic cleavage of the fusion protein, in the inflammatory response and a role for the SH and the secreted form of the G protein in establishing pulmonary infection. Knowledge gained from these studies has also provided the opportunity to develop safe, stable, live attenuated virus vaccine candidates.

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Analysis of Respiratory Syncytial Virus F, G, and SH Proteins in Cell Fusion

Cited by 110 publications

References 0 publications

The Human Respiratory Syncytial Virus Matrix Protein Is Required for Maturation of Viral Filaments

The Human Respiratory Syncytial Virus Matrix Protein Is Required for Maturation of Viral Filaments

Cleavage of the human respiratory syncytial virus fusion protein at two distinct sites is required for activation of membrane fusion

Bovine respiratory syncytial virus infection

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