Nuclear assembly of polyomavirus capsids in insect cells expressing the major capsid protein VP1.
Polyomavirus normally assembles in the nucleus of infected mouse cells. Sf9 insect cells expressing the polyomavirus major capsid protein VP1 were examined by electron microscopy. Capsidlike particles of apparently uniform size were found in the nucleus. Immunogold electron microscopy demonstrated abundant VP1 in the cytoplasm which was not assembled into any recognizable higher-order structure. Cytoplasmic VP1 assembled after the cells were treated with the calcium ionophore ionomycin. Purified VP1 aggregates were shown by negative staining and cryoelectron microscopy to consist predominantly of particles similar to the empty T = 7 viral capsid. Thus, polyomavirus VP1 can assemble in vivo into capsids independent of other viral proteins or DNA. Nuclear assembly may result from increased available calcium in this subcellular compartment.
Montross
L
L
Division of Pediatric Oncology, Children's Hospital, Boston, Massachusetts.
Watkins
S
S
Moreland
R B
RB
Mamon
H
H
Caspar
D L
DL
Garcea
R L
RL
D000818:Animals; D016367:Baculoviridae; D002213:Capsid / Q000235:genetics / Q000378:metabolism* / Q000502:physiology* / Q000648:ultrastructure; D036022:Capsid Proteins; D002460:Cell Line; D002467:Cell Nucleus / Q000378:metabolism; D003001:Cloning, Molecular; D003593:Cytoplasm / Q000378:metabolism; D005822:Genetic Vectors; D015759:Ionomycin / Q000494:pharmacology; D046911:Macromolecular Substances; D008854:Microscopy, Electron; D009024:Morphogenesis; D009036:Moths; D011120:Polyomavirus / Q000235:genetics / Q000648:ultrastructure*
D036022:Capsid Proteins; D046911:Macromolecular Substances; D015759:Ionomycin