Pseudorabies virus envelope glycoproteins gp50 and gII are essential for virus penetration, but only gII is involved in membrane fusion.
To investigate the function of the envelope glycoproteins gp50 and gII of pseudorabies virus in the entry of the virus into cells, we used linker insertion mutagenesis to construct mutant viruses that are unable to express these proteins. In contrast to gD mutants of herpes simplex virus, gp50 mutants, isolated from complementing cells, were able to form plaques on noncomplementing cells. However, progeny virus released from these cells was noninfectious, although the virus was able to adsorb to cells. Thus, the virus requires gp50 to penetrate cells but does not require it in order to spread by cell fusion. This finding indicates that fusion of the virus envelope with the cell membrane is not identical to fusion of the cell membranes of infected and uninfected cells. In contrast to the gp50 mutants, the gII mutant was unable to produce plaques on noncomplementing cells. Examination by electron microscopy of cells infected by the gII mutant revealed that enveloped virus particles accumulated between the inner and outer nuclear membranes. Few noninfectious virus particles were released from the cell, and infected cells did not fuse with uninfected cells. These observations indicate that gII is involved in several membrane fusion events, such as (i) fusion of the viral envelope with the cell membrane during penetration, (ii) fusion of enveloped virus particles with the outer nuclear membrane during the release of nucleocapsids into the cytoplasm, and (iii) fusion of the cell membranes of infected and uninfected cells.
Peeters
B
B
Virology Department, Central Veterinary Institute, Lelystad, The Netherlands.
de Wind
N
N
Hooisma
M
M
Wagenaar
F
F
Gielkens
A
A
Moormann
R
R
D000327:Adsorption; D000818:Animals; D001483:Base Sequence; D002460:Cell Line; D003001:Cloning, Molecular; D016679:Genome, Viral; D011558:Herpesvirus 1, Suid / Q000187:drug effects / Q000235:genetics / Q000502:physiology*; D007700:Kinetics; D008561:Membrane Fusion; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D009838:Oligodeoxyribonucleotides; D010957:Plasmids; D011092:Polyethylene Glycols / Q000494:pharmacology; D015183:Restriction Mapping; D013552:Swine; D014759:Viral Envelope Proteins / Q000235:genetics / Q000502:physiology*; D010948:Viral Plaque Assay; D014771:Virion / Q000187:drug effects / Q000235:genetics / Q000502:physiology
D009838:Oligodeoxyribonucleotides; D014759:Viral Envelope Proteins; C042966:pseudorabies virus glycoproteins; D011092:Polyethylene Glycols