Key Laboratory of Molecular Microbiology and Biotechnology (Ministry of Education), College of Life Sciences, Nankai University, Tianjin, China.
Cell Microbiol. 2010 Aug;12(8):1098-107. doi: 10.1111/j.1462-5822.2010.01453.x. Epub 2010 Feb 9.
Microtubules are essential components of the cytoskeleton that participate in a variety of cellular processes such as cell division and migration. In addition, there is a growing body of evidence implicating a role for microtubules in intracellular viral transport. In this study, we found that pharmacological disruption of microtubules remarkably blocked bovine immunodeficiency virus (BIV) movement from the cell periphery to the perinuclear region, a process known as retrograde transport. A similar effect was observed by inhibiting function of the microtubule-associated motor protein dynein. By yeast two-hybrid assay, we found that the capsid protein (CA) of BIV interacted with the dynein light-chain component LC8. Immunoprecipitation and GST-pulldown assays further demonstrated an interaction between CA and LC8 in mammalian cells. In addition, our data revealed LC8 as a linker between BIV particles and microtubules. Retrograde transport of BIV was significantly inhibited by knockdown of LC8 expression. Our findings present the first evidence that incoming BIV particles employ host microtubule/dynein machinery for transport towards the perinuclear region. In addition, our data indicate that the LC8-CA interaction is a potential target for the design of antiviral strategies.
微管是细胞骨架的重要组成部分，参与多种细胞过程，如细胞分裂和迁移。此外，越来越多的证据表明微管在细胞内病毒运输中起作用。在这项研究中，我们发现微管的药理学破坏显著阻止了牛免疫缺陷病毒（BIV）从细胞外周向核周区域的运动，这一过程称为逆行运输。通过抑制微管相关运动蛋白动力蛋白的功能也观察到了类似的效果。通过酵母双杂交试验，我们发现 BIV 的衣壳蛋白（CA）与动力蛋白轻链成分 LC8 相互作用。免疫沉淀和 GST 下拉试验进一步证明了在哺乳动物细胞中 CA 和 LC8 之间的相互作用。此外，我们的数据显示 LC8 是 BIV 颗粒与微管之间的连接物。LC8 表达的敲低显著抑制了 BIV 的逆行运输。我们的研究结果首次表明，进入的 BIV 颗粒利用宿主微管/动力蛋白机制向核周区域运输。此外，我们的数据表明 LC8-CA 相互作用是设计抗病毒策略的潜在靶点。
