Expression and subcellular targeting of canine parvovirus capsid proteins in baculovirus-transduced NLFK cells.
A mammalian baculovirus delivery system was developed to study targeting in Norden Laboratories feline kidney (NLFK) cells of the capsid proteins of canine parvovirus (CPV), VP1 and VP2, or corresponding counterparts fused to EGFP. VP1 and VP2, when expressed alone, both had equal nuclear and cytoplasmic distribution. However, assembled form of VP2 had a predominantly cytoplasmic localization. When VP1 and VP2 were simultaneously present in cells, their nuclear localization increased. Thus, confocal immunofluorescence analysis of cells transduced with the different baculovirus constructs or combinations thereof in the absence or presence of infecting CPV revealed that the VP1 protein is a prerequisite for efficient targeting of VP2 to the nucleus. The baculovirus vectors were functional and the genes of interest efficiently introduced to this CPV susceptible mammalian cell line. Thus, we show evidence that the system could be utilized to study targeting of the CPV capsid proteins.
Gilbert
Leona
L
Department of Biological and Environmental Science, University of Jyväskylä, P.O. Box 35, FIN-40014, Finland. leona.gilbert@cc.jyu.fi
Välilehto
Outi
O
Kirjavainen
Sanna
S
Tikka
Päivi J
PJ
Mellett
Mark
M
Käpylä
Pirjo
P
Oker-Blom
Christian
C
Vuento
Matti
M
D000818:Animals; D016367:Baculoviridae / Q000235:genetics*; D036022:Capsid Proteins / Q000032:analysis / Q000235:genetics / Q000378:metabolism*; D002415:Cats; D002460:Cell Line; D002467:Cell Nucleus / Q000737:chemistry; D004285:Dogs; D049452:Green Fluorescent Proteins / Q000032:analysis / Q000235:genetics; D017993:Parvovirus, Canine / Q000378:metabolism*; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D014161:Transduction, Genetic
D036022:Capsid Proteins; D011993:Recombinant Fusion Proteins; C485184:enhanced green fluorescent protein; D049452:Green Fluorescent Proteins