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Journal of Virology, July 2007, p. 6899-6908, Vol. 81, No. 13
0022-538X/07/$08.00+0     doi:10.1128/JVI.02330-05
Copyright © 2007, American Society for Microbiology. All Rights Reserved.

Microtubule-Mediated and Microtubule-Independent Transport of Adenovirus Type 5 in HEK293 Cells{triangledown}

Carmen Yea,{dagger} Joanna Dembowy,{ddagger} Laura Pacione,1 and Martha Brown1*

Dept. of Medical Genetics and Microbiology, University of Toronto, Toronto, Ontario, Canada1

Received 4 November 2005/ Accepted 11 April 2007

Adenovirus serotypes 2 and 5 are taken into cells by receptor-mediated endocytosis, and following release from endosomes, destabilized virions travel along microtubules to accumulate around the nucleus. The entry process culminates in delivery of the viral genome through nuclear pores. This model is based on studies with conventional cell lines, such as HeLa and HEp-2, but in HEK293 cells, which are routinely used in this laboratory because they are permissive for replication of multiple adenovirus serotypes, a different trafficking pattern has been observed. Nuclei of 293 cells have an irregular shape, with an indented region, and virions directly labeled with carboxyfluorescein accumulate in a cluster within that indented region. The clusters, which form in close proximity to the microtubule organizing center (MTOC) and to the Golgi apparatus, are remarkably stable; a fluorescent signal can be seen in the MTOC region up to 16 h postinfection. Furthermore, if cells are infected and then undergo mitosis after the cluster is formed, the signal is found at each spindle pole. Despite the sequestration of virions near the MTOC, 293 cells are no less sensitive than other cells to productive infection with adenovirus. Even though cluster formation depends on intact microtubules, infectivity is not compromised by disruption of microtubules with either nocodazole or colchicine, as determined by expression of an enhanced green fluorescent protein reporter gene inserted in the viral genome. These results indicate that virion clusters do not represent the infectious pathway and suggest an alternative route to the nucleus that does not depend on nocodazole-sensitive microtubules.

* Corresponding author. Mailing address: Dept. of Medical Genetics and Microbiology, University of Toronto, Medical Sciences Building, 1 King's College Circle, Toronto, Ontario, Canada M5S 1A8. Phone: (416) 978-5853. Fax: (416) 978-6885. E-mail: martha.brown{at}utoronto.ca

{triangledown} Published ahead of print on 18 April 2007.

{dagger} Present address: Division of Microbiology, The Hospital for Sick Children, Toronto, Ontario, Canada.

{ddagger} Present address: Samuel Lunenfeld Research Institute, Mount Sinai Hospital, Toronto, Ontario, Canada.

Journal of Virology, July 2007, p. 6899-6908, Vol. 81, No. 13
0022-538X/07/$08.00+0     doi:10.1128/JVI.02330-05
Copyright © 2007, American Society for Microbiology. All Rights Reserved.



This article has been cited by other articles:

  • Russell, W. C. (2009). Adenoviruses: update on structure and function. J. Gen. Virol. 90: 1-20 [Abstract] [Full Text]  


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