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Journal of Virology, December 1999, p. 10371-10376, Vol. 73, No. 12
0022-538X/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.

Dynamin Is Required for Recombinant Adeno-Associated Virus Type 2 Infection

Dongsheng Duan,1,2 Qiang Li,1 Aimee W. Kao,3 Yongping Yue,1 Jeffrey E. Pessin,3 and John F. Engelhardt1,2,4,*

Department of Anatomy and Cell Biology,1 Department of Internal Medicine,4 Center for Gene Therapy,2 and Department of Physiology and Biophysics,3 College of Medicine, The University of Iowa, Iowa City, Iowa 52242

Received 1 June 1999/Accepted 3 September 1999

Recombinant adeno-associated virus (rAAV) vectors for gene therapy of inherited disorders have demonstrated considerable potential for molecular medicine. Recent identification of the viral receptor and coreceptors for AAV type 2 (AAV-2) has begun to explain why certain organs may demonstrate higher efficiencies of gene transfer with this vector. However, the mechanisms by which AAV-2 enters cells remain unknown. In the present report, we have examined whether the endocytic pathways of rAAV-2 are dependent on dynamin, a GTPase protein involved in clathrin-mediated internalization of receptors and their ligands from the plasma membrane. Using a recombinant adenovirus expressing a dominant-inhibitory form of dynamin I (K44A), we have demonstrated that rAAV-2 infection is partially dependent on dynamin function. Overexpression of mutant dynamin I significantly inhibited AAV-2 internalization and gene delivery, but not viral binding. Furthermore, colocalization of rAAV and transferrin in the same endosomal compartment provides additional evidence that clathrin-coated pits are the predominant pathway for endocytosis of AAV-2 in HeLa cells.

* Corresponding author. Mailing address: Department of Anatomy and Cell Biology, University of Iowa, College of Medicine, 51 Newton Rd., Room 1-111 BSB, Iowa City, IA 52242-1109. Phone: (319) 335-7753. Fax: (319) 335-7198. E-mail: john-engelhardt{at}uiowa.edu.

Journal of Virology, December 1999, p. 10371-10376, Vol. 73, No. 12
0022-538X/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.


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