title: Glycoproteins M and N of pseudorabies virus form a disulfide-linked complex.
abstract: Genes homologous to the herpes simplex virus UL49.5 open reading frame are conserved throughout the Herpesviridae. In the alphaherpesvirus pseudorabies virus (PrV), the UL49.5 product is an O-glycosylated structural protein of the viral envelope, glycoprotein N (gN) (A. Jöns, H. Granzow, R. Kuchling, and T. C. Mettenleiter, J. Virol. 70:1237-1241, 1996). For functional characterization of gN, a gN-negative PrV mutant, PrV-gNbeta, and the corresponding rescuant, PrV-gNbetaR, were constructed, gN-negative PrV was able to productively replicate on noncomplementing cells, and one-step growth in cell culture was only slightly reduced compared to that of wild-type PrV. However, penetration was significantly delayed. In indirect immunofluorescence assays with rabbit serum directed against baculovirus-expressed gN, specific staining of wild-type PrV-infected cells occurred only after permeabilization of cells, whereas live cells failed to react with the antiserum. This indicates the lack of surface accessibility of gN in the plasma membrane of a PrV-infected cell. Western blot analyses and radioimmunoprecipitation experiments under reducing and nonreducing conditions led to the discovery of a heteromeric complex composed of gM and gN. The complex was stable in the absence of 2-mercaptoethanol but dissociated after the addition of the reducing agent, indicating that the partners are linked by disulfide bonds. Finally, gN was absent from gM-negative PrV virions, whereas gM was readily detected in virions in the absence of gN. Thus, gM appears to be required for virion localization of gN.
authors.affiliation: Institute of Molecular and Cellular Virology, Friedrich-Loeffler-Institutes, Federal Research Centre for Virus Diseases of Animals, Insel Riems, Germany.
authors.forename: A
authors.initials: A
authors.lastname: Jöns
authors.forename: J M
authors.initials: JM
authors.lastname: Dijkstra
authors.forename: T C
authors.initials: TC
authors.lastname: Mettenleiter
mesh_terms: D000818:Animals; D001483:Base Sequence; D002417:Cattle; D002460:Cell Line; D017931:DNA Primers / Q000235:genetics; D004220:Disulfides / Q000737:chemistry; D016679:Genome, Viral; D011558:Herpesvirus 1, Suid / Q000235:genetics / Q000378:metabolism* / Q000502:physiology; D046911:Macromolecular Substances; D008970:Molecular Weight; D009154:Mutation; D011817:Rabbits; D014759:Viral Envelope Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D014779:Virus Replication
chemical_list: D017931:DNA Primers; D004220:Disulfides; D046911:Macromolecular Substances; D014759:Viral Envelope Proteins; C097733:glycoprotein N, Pseudorabies virus; C042966:pseudorabies virus glycoproteins