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Journal of Virology, March 2005, p. 2720-2728, Vol. 79, No. 5
0022-538X/05/$08.00+0     doi:10.1128/JVI.79.5.2720-2728.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.

Improvement in Nuclear Entry and Transgene Expression of Baculoviruses by Disintegration of Microtubules in Human Hepatocytes

Mirka Salminen,1 Kari J. Airenne,2 Rita Rinnankoski,1 Jenni Reimari,1 Outi Välilehto,1 Johanna Rinne,1 Sanna Suikkanen,1 Sari Kukkonen,2 Seppo Ylä-Herttuala,2 Markku S. Kulomaa,1 and Maija Vihinen-Ranta1*

NanoScience Center, Department of Biological and Environmental Science, University of Jyväskylä, Jyväskylä,1 AI Virtanen Institute, Department of Biotechnology and Molecular Medicine, University of Kuopio, Kuopio, Finland2

Received 10 August 2004/ Accepted 14 September 2004

Autographa californica multicapsid nucleopolyhedrovirus (AcMNPV), a potent virus for mammalian cell gene delivery, possesses an ability to transduce mammalian cells without viral replication. We examined the role of the cellular cytoskeleton in the cytoplasmic trafficking of viral particles toward the nucleus in human hepatic cells. Microscopic studies showed that capsids were found in the nucleus after either viral inoculation or cytoplasmic microinjection of nucleocapsids. The presence of microtubule (MT) depolymerizing agents caused the amount of nuclear capsids to increase. Overexpression of p50/dynamitin, an inhibitor of dynein-dependent endocytic trafficking from peripheral endosomes along MTs toward late endosomes, did not significantly affect the amount of nuclear accumulation of nucleocapsids in the inoculated cells, suggesting that viral nucleocapsids are released into the cytosol during the early stages of the endocytic pathway. Moreover, studies with recombinant viruses containing the nuclear-targeted expression ß-galactosidase gene (ß-gal) showed a markedly increased level in the cellular expression of ß-galactosidase in the presence of MT-disintegrating drugs. The maximal increase in expression at 10 h postinoculation was observed in the presence of 80 µM nocodazole or 10 µM vinblastine. Together, these data suggest that the intact MTs constitute a barrier to baculovirus transport toward the nucleus.

* Corresponding author. Mailing address: Department of Biological and Environmental Science, P.O. Box 35 (YAB), FIN-40014 University of Jyväskylä, Jyväskylä, Finland. Phone: (358) 14-2604209. Fax: (358) 14-2602221. E-mail: mvihinen{at}jyu.fi.

Journal of Virology, March 2005, p. 2720-2728, Vol. 79, No. 5
0022-538X/05/$08.00+0     doi:10.1128/JVI.79.5.2720-2728.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.



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  • Matilainen, H., Rinne, J., Gilbert, L., Marjomaki, V., Reunanen, H., Oker-Blom, C. (2005). Baculovirus Entry into Human Hepatoma Cells. J. Virol. 79: 15452-15459 [Abstract] [Full Text]  


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