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Journal of Virology, October 2003, p. 10270-10279, Vol. 77, No. 19
0022-538X/03/$08.00+0     DOI: 10.1128/JVI.77.19.10270-10279.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.

Exploitation of Microtubule Cytoskeleton and Dynein during Parvoviral Traffic toward the Nucleus

Sanna Suikkanen, Tuula Aaltonen, Marjukka Nevalainen, Outi Välilehto, Laura Lindholm, Matti Vuento, and Maija Vihinen-Ranta*

Department of Biological and Environmental Science, University of Jyväskylä, FIN-40500 Jyvaskyla, Finland

Received 21 April 2003/ Accepted 30 June 2003

Canine parvovirus (CPV), a model virus for the study of parvoviral entry, enters host cells by receptor-mediated endocytosis, escapes from endosomal vesicles to the cytosol, and then replicates in the nucleus. We examined the role of the microtubule (MT)-mediated cytoplasmic trafficking of viral particles toward the nucleus. Immunofluorescence and immunoelectron microscopy showed that capsids were transported through the cytoplasm into the nucleus after cytoplasmic microinjection but that in the presence of MT-depolymerizing agents, viral capsids were unable to reach the nucleus. The nuclear accumulation of capsids was also reduced by microinjection of an anti-dynein antibody. Moreover, electron microscopy and light microscopy experiments demonstrated that viral capsids associate with tubulin and dynein in vitro. Coprecipitation studies indicated that viral capsids interact with dynein. When the cytoplasmic transport process was studied in living cells by microinjecting fluorescently labeled capsids into the cytoplasm of cells containing fluorescent tubulin, capsids were found in close contact with MTs. These results suggest that intact MTs and the motor protein dynein are required for the cytoplasmic transport of CPV capsids and contribute to the accumulation of the capsid in the nucleus.

* Corresponding author. Mailing address: Department of Biological and Environmental Science, P.O. Box 35, University of Jyväskylä, Survontie 9, FIN-40500 Jyvaskyla, Finland. Phone: (358) 14 2604209. Fax: (358) 14 2602271. E-mail: mvihinen{at}jyu.fi.

Journal of Virology, October 2003, p. 10270-10279, Vol. 77, No. 19
0022-538X/03/$08.00+0     DOI: 10.1128/JVI.77.19.10270-10279.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.



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