Baculovirus infection of nondividing mammalian cells: mechanisms of entry and nuclear transport of capsids.
We have studied the infection pathway of Autographa californica multinuclear polyhedrosis virus (baculovirus) in mammalian cells. By titration with a baculovirus containing a green fluorescent protein cassette, we found that several, but not all, mammalian cell types can be infected efficiently. In contrast to previous suggestions, our data show that the asialoglycoprotein receptor is not required for efficient infection. We demonstrate for the first time that this baculovirus can infect nondividing mammalian cells, which implies that the baculovirus is able to transport its genome across the nuclear membrane of mammalian cells. Our data further show that the virus enters via endocytosis, followed by an acid-induced fusion event, which releases the nucleocapsid into the cytoplasm. Cytochalasin D strongly reduces the infection efficiency but not the delivery of nucleocapsids to the cytoplasm, suggesting involvement of actin filaments in cytoplasmic transport of the capsids. Electron microscopic analysis shows the cigar-shaped nucleocapsids located at nuclear pores of nondividing cells. Under these conditions, we observed the viral genome, major capsid protein, and electron-dense capsids inside the nucleus. This suggests that the nucleocapsid is transported through the nuclear pore. This mode of transport seems different from viruses with large spherical capsids, such as herpes simplex virus and adenovirus, which are disassembled before nuclear transport of the genome. The implications for the application of baculovirus or its capsid proteins in gene therapy are discussed.
van Loo
N D
ND
Department of Cell Biology, Erasmus University, 3000 DR Rotterdam, The Netherlands.
Fortunati
E
E
Ehlert
E
E
Rabelink
M
M
Grosveld
F
F
Scholte
B J
BJ
D000818:Animals; D002460:Cell Line; D002467:Cell Nucleus / Q000821:virology*; D004705:Endocytosis; D011992:Endosomes / Q000821:virology; D005455:Fluorescent Antibody Technique; D016679:Genome, Viral; D049452:Green Fluorescent Proteins; D006801:Humans; D006863:Hydrogen-Ion Concentration; D017404:In Situ Hybridization, Fluorescence; D008164:Luminescent Proteins / Q000235:genetics / Q000378:metabolism; D008854:Microscopy, Electron; D022022:Nuclear Pore / Q000378:metabolism / Q000821:virology; D019251:Nucleocapsid / Q000378:metabolism*; D017924:Nucleopolyhedroviruses / Q000235:genetics / Q000502:physiology*; D051381:Rats; D011991:Receptors, Virus / Q000502:physiology; D016196:S Phase / Q000502:physiology; D014407:Tumor Cells, Cultured
D008164:Luminescent Proteins; D011991:Receptors, Virus; D049452:Green Fluorescent Proteins