Analysis of respiratory syncytial virus F, G, and SH proteins in cell fusion.
Recombinant expression of the human respiratory syncytial virus (RSV) fusion (F) glycoprotein, receptor-binding glycoprotein (G), and small hydrophobic (SH) protein was performed to determine the role(s) of these proteins in syncytia formation. These studies used a vaccinia virus expressing the bacteriophage (T7) RNA polymerase gene and plasmid vectors containing the RSV genes under the control of a T7 promoter. Within the context of this expression system, expression of any individual RSV gene, or coexpression of F+G genes, did not elicit the formation of syncytia. However, at plasmid input levels which were 10-fold higher than those normally used, coexpression of F+G induced low but detectable levels of cell fusion. In contrast, coexpression of F, G, and SH together elicited extensive cell fusion resembling that of an authentically infected cell monolayer. In addition, coexpression of F and SH elicited significant cell fusion, although to a lesser extent than was observed when G was included. Cell fusion induced by coexpression of F+SH was found to be specific to the RSV proteins, since coexpression of SH with the analogous F proteins from human parainfluenza virus type 3, human parainfluenza virus type 2, Sendai virus, or simian virus type 5 (SV5) did not elicit cell fusion. Finally, coexpression of the SV5 SH protein with the RSV or SV5 glycoproteins also failed to induce syncytia, suggesting type-specific restrictions between the two sets of viral proteins.
Heminway
B R
BR
Department of Molecular Biology, Cleveland Clinic Foundation, Ohio 44195.
Yu
Y
Y
Tanaka
Y
Y
Perrine
K G
KG
Gustafson
E
E
Bernstein
J M
JM
Galinski
M S
MS
D000818:Animals; D001483:Base Sequence; D002459:Cell Fusion; D015707:HN Protein; D008969:Molecular Sequence Data; D011994:Recombinant Proteins; D012136:Respiratory Syncytial Viruses / Q000502:physiology*; D014162:Transfection; D014616:Vaccinia virus / Q000235:genetics; D014759:Viral Envelope Proteins; D014760:Viral Fusion Proteins / Q000502:physiology*; D014764:Viral Proteins / Q000502:physiology*
D015707:HN Protein; D011994:Recombinant Proteins; D014759:Viral Envelope Proteins; D014760:Viral Fusion Proteins; D014764:Viral Proteins; C116204:attachment protein G