Dynamin is required for recombinant adeno-associated virus type 2 infection.
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.
Duan
D
D
Department of Anatomy, College of Medicine, The University of Iowa, Iowa City, Iowa 52242, USA.
Li
Q
Q
Kao
A W
AW
Yue
Y
Y
Pessin
J E
JE
Engelhardt
J F
JF
D002461:Cell Line, Transformed; D000229:Dependovirus / Q000378:metabolism / Q000502:physiology*; D034284:Dynamin I; D034281:Dynamins; D004705:Endocytosis; D020558:GTP Phosphohydrolases / Q000096:biosynthesis / Q000235:genetics / Q000378:metabolism*; D006367:HeLa Cells; D006801:Humans; D016296:Mutagenesis; D011995:Recombination, Genetic; D014168:Transferrin / Q000378:metabolism
D014168:Transferrin; D034284:Dynamin I; D020558:GTP Phosphohydrolases; D034281:Dynamins