Simultaneous deletion of pseudorabies virus tegument protein UL11 and glycoprotein M severely impairs secondary envelopment.
The pseudorabies virus (PrV) proteins UL11, glycoprotein E (gE), and gM are involved in secondary envelopment of tegumented nucleocapsids in the cytoplasm. To assess the relative contributions of these proteins to the envelopment process, virus mutants with deletions of either UL11, gM, or gE as well as two newly constructed mutant viruses with simultaneous deletions of UL11 and gE or of UL11 and gM were analyzed in cell culture for their growth phenotype. We show here that simultaneous deletion of UL11 and gE reduced plaque size in an additive manner over the reduction observed by deletion of only UL11 or gE. However, one-step growth was not further impaired beyond the level of the UL11 deletion mutant. Moreover, in electron microscopic analyses PrV-DeltaUL11/gE exhibited a phenotype similar to that of the UL11 mutant virus. In contrast, plaque formation was virtually abolished by the simultaneous absence of UL11 and gM, and one-step growth was significantly reduced. Electron microscopy showed the presence of huge intracytoplasmic inclusions in PrV-DeltaUL11/gM-infected cells, with a size reaching 3 micro m and containing nucleocapsids embedded in tegument. We hypothesize that UL11 and gM are involved in different steps during secondary envelopment and that simultaneous deletion of both interrupts both processes, resulting in the observed drastic impairment of secondary envelopment.
Kopp
Martina
M
Institutes of Molecular Biology. Infectology, Friedrich-Loeffler-Institutes, Federal Research Centre for Virus Diseases of Animals, D-17493 Greifswald-Insel Riems, Germany.
Granzow
Harald
H
Fuchs
Walter
W
Klupp
Barbara
B
Mettenleiter
Thomas C
TC
D000818:Animals; D002460:Cell Line; D017353:Gene Deletion; D011558:Herpesvirus 1, Suid / Q000254:growth & development*; D016253:Microscopy, Immunoelectron; D011817:Rabbits; D014759:Viral Envelope Proteins / Q000235:genetics* / Q000378:metabolism; D010948:Viral Plaque Assay; D015678:Viral Structural Proteins / Q000235:genetics* / Q000378:metabolism; D014771:Virion / Q000378:metabolism
D014759:Viral Envelope Proteins; D015678:Viral Structural Proteins; C042966:pseudorabies virus glycoproteins