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Journal of Virology, March 2002, p. 2340-2349, Vol. 76, No. 5
0022-538X/02/$04.00+0     DOI: 10.1128/jvi.76.5.2340-2349.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.

Endocytosis of Adeno-Associated Virus Type 5 Leads to Accumulation of Virus Particles in the Golgi Compartment

*** Ursula Bantel-Schaal,1* Birgit Hub,1 and Juergen Kartenbeck2

Forschungsschwerpunkt Angewandte Tumorvirologie F0400, Abteilung Pathogenitätsmechanismen,1 Forschungsschwerpunkt Krebsentstehung und Differenzierung A0100, Abteilung für Zellbiologie, Deutsches Krebsforschungszentrum Heidelberg, 69120 Heidelberg, Germany2

Received 7 November 2001/ Accepted 19 November 2001

Among the adeno-associated virus (AAV) serotypes which are discussed as vectors for gene therapy AAV type 5 (AAV5) represents a candidate with unique advantages. To further our knowledge on AAV5-specific characteristics, we studied the entry pathway of wild-type virus in HeLa cells in the absence of helper virus by immunofluorescence and electron microscopy and by Western blot analysis. We found virus binding at the apical cell surface, especially at microvilli and, with increasing incubation time, virus accumulation at cell-cell boundaries. The different binding kinetics suggest different binding properties at apical versus lateral plasma membranes. Endocytosis of viruses was predominantly by clathrin-coated vesicles from both membrane domains; however, particles were also detected in noncoated pits. AAV5 particles were mainly routed to the Golgi area, where they could be detected within cisternae of the trans-Golgi network and within vesicles associated with cisternae and with the dictyosomal stacks of the Golgi apparatus. These data suggest that AAV5 makes use of endocytic routes that have hitherto not been described as pathways for virus entry.

* Corresponding author. Mailing address: Forschungsschwerpunkt Angewandte Tumorvirologie F0400, Abteilung Pathogenitätsmechanismen, Deutsches Krebsforschungszentrum Heidelberg, Im Neuenheimer Feld 242, 69120 Heidelberg, Germany. Phone: 49-6221-424823. Fax: 49-6221-424902. E-mail: u.bantel-schaal{at}dkfz-heidelberg.de.

Journal of Virology, March 2002, p. 2340-2349, Vol. 76, No. 5
0022-538X/02/$04.00+0     DOI: 10.1128/jvi.76.5.2340-2349.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.



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