PMID:11369882
| Citation |
Sugrue, RJ, Brown, C, Brown, G, Aitken, J and McL Rixon, HW (2001) Furin cleavage of the respiratory syncytial virus fusion protein is not a requirement for its transport to the surface of virus-infected cells. J. Gen. Virol. 82:1375-86 |
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| Abstract |
The intracellular cleavage of respiratory syncytial virus (RSV) fusion (F) protein by furin was examined. In RSV-infected LoVo cells, which express an inactive form of furin, and in RSV-infected Vero cells treated with the furin inhibitor decanoyl-Arg-Val-Lys-Arg-chloromethyl ketone (dec-RVKR-cmk), the F protein was expressed as a non-cleaved 73 kDa species. In both cases the F protein was initially expressed as an endoglycosidase H (Endo H)-sensitive precursor (F0(EHs)) which was modified approximately 40 min post-synthesis by the addition of complex carbohydrates to produce the Endo H-resistant form (F0(EHr)). The size and glycosylation state of F0(EHr) were identical to a transient intermediate form of non-cleaved F protein which was detected in RSV-infected Vero cells in the absence of inhibitor. Cell surface biotinylation and surface immunofluorescence staining showed that F0(EHr) was present on the surface of RSV-infected cells. RSV filaments have been shown to be the predominant form of the budding virus that is detected during virus replication. Analysis of the RSV-infected cells using scanning electron microscopy (SEM) showed that, in the presence of dec-RVKR-cmk, virus budding was impaired, producing fewer and much smaller viral filaments than in untreated cells. A comparison of immunofluorescence and SEM data showed that F0(EHr) was routed to the surface of virus-infected cells but not located in these smaller structures. Our findings suggest that activation of the F protein is required for the efficient formation of RSV filaments. |
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| Keywords |
Animals; Biotinylation; Cell Line; Cell Membrane/metabolism; Cell Membrane/ultrastructure; Cell Membrane/virology; Cercopithecus aethiops; Fluorescent Antibody Technique; Furin; Glycosylation; Kinetics; Microscopy, Electron, Scanning; Molecular Weight; Mutation/genetics; Protein Processing, Post-Translational/drug effects; Protein Transport/drug effects; Respiratory Syncytial Viruses/drug effects; Respiratory Syncytial Viruses/growth & development; Respiratory Syncytial Viruses/metabolism; Respiratory Syncytial Viruses/ultrastructure; Serine Proteinase Inhibitors/pharmacology; Subtilisins/antagonists & inhibitors; Subtilisins/genetics; Subtilisins/metabolism; Vero Cells; Viral Fusion Proteins/chemistry; Viral Fusion Proteins/metabolism; Viral Fusion Proteins/ultrastructure; Virus Assembly/drug effects |
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