Equine herpesvirus 1 (EHV-1) glycoprotein M: effect of deletions of transmembrane domains.
Equine herpesvirus 1 (EHV-1) recombinants that carry either a deletion of glycoprotein M (gM) or express mutant forms of gM were constructed. The recombinants were derived from strain Kentucky A (KyA), which also lacks genes encoding gE and gI. Plaques on RK13 cells induced by the gM-negative KyA were reduced in size by 80%, but plaque sizes were restored to wild-type levels on gM-expressing cells. Electron microscopic studies revealed a massive defect in virus release after the deletion of gM in the gE- and gI-negative KyA, which was caused by a block in secondary envelopment of virions at Golgi vesicles. Recombinant KyA expressing mutant gM with deletions of predicted transmembrane domains was generated and characterized. It was shown that mutant gM was expressed and formed dimeric and oligomeric structures. However, subcellular localization of mutant gM proteins differed from that of wild-type gM. Mutant glycoproteins were not transported to the Golgi network and consequently were not incorporated into the envelope of extracellular virions. Also, a small plaque phenotype of mutant viruses that was indistinguishable from that of the gM-negative KyA was observed. Plaque sizes of mutant viruses were restored to wild-type levels by plating onto RK13 cells constitutively expressing full-length EHV-1 gM, indicating that mutant proteins did not exert a transdominant negative effect on wild-type gM.
Seyboldt
C
C
Institutes of Molecular Biology, Insel Riems, D-17498, Germany.
Granzow
H
H
Osterrieder
N
N
D000595:Amino Acid Sequence; D000818:Animals; D002460:Cell Line; D017931:DNA Primers; D004861:Herpesvirus 1, Equid / Q000235:genetics / Q000502:physiology* / Q000648:ultrastructure; D006736:Horses; D007700:Kinetics; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D017384:Sequence Deletion; D014759:Viral Envelope Proteins / Q000737:chemistry / Q000235:genetics*; D010948:Viral Plaque Assay
D017931:DNA Primers; D014759:Viral Envelope Proteins; C418522:glycoprotein M, Equid herpesvirus 1