Effect of inhibition of dynein function and microtubule-altering drugs on AAV2 transduction.
Over the past decade, adeno-associated (AAV) virus has emerged as an important vector for gene therapy. As a result, understanding its basic biology, including intracellular trafficking, has become increasingly important. Here, we describe the effect of inhibiting dynein function or altering the state of microtubule polymerization on rAAV2 transduction. Overexpression of dynamitin, resulting in a functional inhibition of the minus-end-directed microtubule motor protein dynein, did not inhibit transduction. Equally, treatment of cells with nocodazole, or concentrations of vinblastine that result in the disruption of microtubules, had no significant effect on transduction. In contrast, high concentrations of Taxol and vinblastine, resulting in microtubule stabilization and the formation of tubulin paracrystals respectively, reduced rAAV2 transduction in a vector-dose-dependent manner. These results demonstrate that AAV2 can infect HeLa cells independently of dynein function or an intact microtubule network.
Hirosue
Sachiko
S
Department of Gene and Cell Medicine, Mount Sinai School of Medicine, 1 Gustave L. Levy Place, Box 1496, New York, NY 10029-6514, USA.
Senn
Karin
K
Clément
Nathalie
N
Nonnenmacher
Mathieu
M
Gigout
Laure
L
Linden
R Michael
RM
Weber
Thomas
T
D000970:Antineoplastic Agents / Q000494:pharmacology; D000229:Dependovirus / Q000235:genetics / Q000378:metabolism / Q000472:pathogenicity*; D004398:Dyneins / Q000037:antagonists & inhibitors* / Q000378:metabolism; D006367:HeLa Cells; D006801:Humans; D008870:Microtubules / Q000187:drug effects* / Q000378:metabolism*; D015739:Nocodazole / Q000494:pharmacology; D017239:Paclitaxel / Q000494:pharmacology; D011995:Recombination, Genetic; D014161:Transduction, Genetic; D050257:Tubulin Modulators / Q000494:pharmacology; D014747:Vinblastine / Q000494:pharmacology
D000970:Antineoplastic Agents; D050257:Tubulin Modulators; D014747:Vinblastine; D004398:Dyneins; D017239:Paclitaxel; D015739:Nocodazole