Hirosue Sachiko, Senn Karin, Clément Nathalie, Nonnenmacher Mathieu, Gigout Laure, Linden R Michael, Weber Thomas
Department of Gene and Cell Medicine, Mount Sinai School of Medicine, 1 Gustave L. Levy Place, Box 1496, New York, NY 10029-6514, USA.
Virology. 2007 Oct 10;367(1):10-8. doi: 10.1016/j.virol.2007.05.009. Epub 2007 Jun 22.
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.
在过去十年中，腺相关（AAV）病毒已成为基因治疗的重要载体。因此，了解其基本生物学特性，包括细胞内运输，变得越来越重要。在这里，我们描述了抑制动力蛋白功能或改变微管聚合状态对rAAV2转导的影响。过表达发动蛋白，导致对负端定向微管运动蛋白动力蛋白的功能抑制，并未抑制转导。同样，用诺考达唑或导致微管破坏的长春碱浓度处理细胞，对转导没有显著影响。相反，高浓度的紫杉醇和长春碱分别导致微管稳定和微管蛋白副晶体的形成，以载体剂量依赖的方式降低了rAAV2转导。这些结果表明，AAV2可以独立于动力蛋白功能或完整的微管网络感染HeLa细胞。
