Expression of the polyomavirus VP2 and VP3 proteins in insect cells: coexpression with the major capsid protein VP1 alters VP2/VP3 subcellular localization.
During polyomavirus infection the capsid proteins are synthesized in the cytoplasm and transported into the nucleus were virion assembly occurs. Expression of the major capsid protein VP1 in Sf9 insect cells results in the accumulation of capsid-like particles in the nucleus, independent of the presence of the minor capsid proteins VP2 and VP3 or the viral DNA (Montross et al., J. Virol. 65, 4991-4998, 1991). Sf9 cells infected with baculovirus vectors expressing the polyomavirus minor capsid proteins VP2 and VP3 were examined. VP2 was myristylated in Sf9 cells, as seen during polyomavirus infection of mouse cells. Immunoprecipitation of lysates from co-infected cells demonstrated an association between VP1 and VP2. As determined by immunogold electron microscopy, when expressed alone VP2 was associated with membrane structures in the cytoplasm and VP3 was diffusely localized in the cytoplasm. When co-infected with a VP1 expressing baculovirus, both VP2 and VP3 became predominantly localized to the nucleus in association with capsid-like structures. Thus, the polyomavirus capsid proteins interact in vivo and alter their subcellular localization as a consequence.
Division of Pediatric Oncology, Dana-Farber Cancer Institute, Boston, Massachusetts 02115.
S E
SE
Delos
L
L
Montross
R B
RB
Moreland
R L
RL
Garcea
D000818:Animals; D002213:Capsid / Q000096:biosynthesis* / Q000235:genetics; D036022:Capsid Proteins; D002451:Cell Compartmentation; D002467:Cell Nucleus / Q000737:chemistry; D002478:Cells, Cultured; D003593:Cytoplasm / Q000737:chemistry; D015870:Gene Expression; D007150:Immunohistochemistry; D008566:Membranes / Q000737:chemistry; D009036:Moths; D019814:Myristic Acid; D009227:Myristic Acids / Q000378:metabolism; D011120:Polyomavirus / Q000235:genetics* / Q000648:ultrastructure; D011499:Protein Processing, Post-Translational
D036022:Capsid Proteins; D009227:Myristic Acids; C442543:VP2 protein, Polyomavirus; D019814:Myristic Acid