Analysis of axonal transport of herpes simplex virus and examination of the possibility as a virus vector

有微管依赖的马达蛋白动力蛋白逆行轴突运输，驱动蛋白顺行运输。我们研究了这些蛋白与病毒膜蛋白的相互作用，以探讨单纯疱疹病毒（HSV）轴突转运的机制。该研究表明，间层蛋白UL 34对神经元特异性运动蛋白动力蛋白IC 1A变体具有高亲和力，对常规动力蛋白IC 2C变体和货物衔接蛋白DCTN 1（p150^）具有低亲和力<GIued>。为了研究体内相互作用，制备表达GFP-UL 34融合蛋白的重组病毒（rGFP-UL 34）。rGFP-UL 34感染HeLa细胞后，UL 34定位于细胞核，聚集于核膜，但不聚集于微管。不幸的是，UL 34表达的辅助细胞对于rGFP-UL 34的复制是必需的。这一结果表明GFP-UL 34融合蛋白损害了片段蛋白的物理功能和/或不包含在病毒体中。 ...更多信息 rt证明了US 11和作为另一种马达蛋白的普遍存在的驱动蛋白之间的相互作用。我们制备了表达GFP-US 11融合蛋白的重组病毒（rGFP-US 11），以观察HSV的顺行轴突运输。在rGFP-US 11感染的HeLa细胞中，US 11表达于细胞质中，定位于核仁而非微管。为了研究病毒蛋白激酶（UL 13和US 3）对轴突运输的参与，制备了以下重组病毒：rGFP-US 11病毒、US 3缺失病毒、UL 13缺失病毒、US 3缺失/rGFP-US 11病毒、UL 13缺失/rGFP-US 11病毒。不幸的是，这些病毒感染的细胞之间的差异没有观察到病毒释放到培养基中。由于US 11对于HSV在体内的轴突运输是必需的，但对于培养细胞中的生产性病毒生长不是必需的，因此我们正在研究体内实验，例如神经节细胞的原代培养或啮齿动物的体内系统。另一方面，我们研究了病毒基因组的包装，特别是单位长度基因组形成的切割机制。少

There are the microtubule-dependent motor protein dynein for retrograde axonal transport, the kinesin for anterograde transport. We were examined the interaction between those proteins and virus tegment proteins for the mechanism of axonal transport of herpes simplex virus (HSV). This examination suggested that the tegument protein UL34 has a high affinity for neuron-specific motor protein dynein IC1A variant, a low affinity for conventional dynein IC2C variant and cargo-adaptor protein DCTN1 (p150^<GIued>). To investigate the interactions In vivo, recombinant virus (rGFP-UL34) expressing a GFP-UL34 fusion protein was prepared. UL34 was located to nucleus and accumulated to nuclear membrane but not to microtubules in rGFP-UL34 infected-HeLa cells. Unfortunately, a UL34-expressed helper cells is necessary for the replication of rGFP-UL34. This result suggests that a GFP-UL34 fusion protein impaired a physical function in tegment protein and/or does not included in a virion.A recent repo … More rt demonstrates an interaction between US11 and the ubiquitous kinesin as the other motor protein. We prepared the recombinant virus (rGFP-US11) expressing a GFP-US11 fusion protein to observe the anterograde axonal transport of HSV. US11 was expressed in the cytoplasm and located to nucleoli but not to microtubules in rGFP-US11 infected-HeLa cells. To investigate the participation of virus protein kinases (UL13 and US3) to the axonal transport, the following recombinant viruses was prepared ; rGFP-US11 virus, US3 deletion virus, UL13 deletion virus, US3 deletion/rGFP-US11 virus, UL13 deletion/rGFP-US11l virus. Unfortunately, the differences among these viruses-infected cells were not observed in the release of viruses to culture media. Since US11 is essential for axonal transport of HSV in vivo but not for productive viral growth in cultured cells, we are studying in vivo experiments such as the primary culture of ganglia cells or in vivo system of rodents. On the other hand, we investigate the packaging of virus genomes, especially cleavage mechanism for the formation of unit-length genomes. Less