Intracellular transport and stability of varicella-zoster virus glycoprotein K.
VZV gK, an essential glycoprotein that is conserved among the alphaherpesviruses, is believed to participate in membrane fusion and cytoplasmic virion morphogenesis based on analogy to its HSV-1 homolog. However, the production of VZV gK-specific antibodies has proven difficult presumably due to its highly hydrophobic nature and, therefore, VZV gK has received limited study. To overcome this obstacle, we inserted a FLAG epitope into gK near its amino terminus and produced VZV recombinants expressing epitope-tagged gK (VZV gK-F). These recombinants grew indistinguishably from native VZV, and FLAG-tagged gK could be readily detected in VZV gK-F-infected cells. FACS analysis established that gK is transported to the plasma membrane of infected cells, while indirect immunofluorescence demonstrated that gK accumulates predominately in the Golgi. Using VZV gK-F-infected cells we demonstrated that VZV gK, like several other herpesvirus glycoproteins, is efficiently endocytosed from the plasma membrane. However, pulse-labeling experiments revealed that the half-life of gK is considerably shorter than that of other VZV glycoproteins including gB, gE and gH. This finding suggests that gK may be required in lower abundance than other viral glycoproteins during virion morphogenesis or viral entry.
Hall
Susan L
SL
Division of Infectious Diseases and Immunology, Saint Louis University School of Medicine, Saint Louis, Missouri 63110-0250, USA.
Govero
Jennifer L
JL
Heineman
Thomas C
TC
D000818:Animals; D001692:Biological Transport; D002460:Cell Line; D002462:Cell Membrane / Q000378:metabolism*; D006023:Glycoproteins / Q000378:metabolism*; D006056:Golgi Apparatus / Q000378:metabolism; D006207:Half-Life; D014645:Herpesvirus 3, Human / Q000502:physiology*; D006801:Humans; D016865:Reassortant Viruses; D011994:Recombinant Proteins / Q000378:metabolism; D014759:Viral Envelope Proteins / Q000378:metabolism*; D014779:Virus Replication
D006023:Glycoproteins; C118859:ORF5 protein, Human herpesvirus 3; D011994:Recombinant Proteins; D014759:Viral Envelope Proteins