Receptor (CD155)-dependent endocytosis of poliovirus and retrograde axonal transport of the endosome
Receptor (CD155)-dependent endocytosis of poliovirus and retrograde axonal transport of the endosome
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DOI：
10.1128/jvi.78.13.7186-7198.2004
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发表时间：
2004-07-01
影响因子：
5.4
通讯作者：
Nomoto, A
中科院分区：
文献类型：
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作者：
Ohka, S;Matsuda, N;Nomoto, A
脊髓灰质炎病毒（PV），当肌肉注射到小牛，被纳入坐骨神经，并导致接种肢体的初步麻痹，在转基因小鼠携带人PV受体（hPVR/CD 155）基因。在这里，我们证明了通过使用免疫电镜PV颗粒存在于囊泡结构的神经末梢的神经肌肉接头。我们还证明，在谷胱甘肽S-转移酶下拉实验中，动力蛋白轻链，Tctex-1，直接与hPVR的细胞质结构域相互作用。在用hPVRs表达载体转染的分化的大鼠PC 12细胞的轴突中，由PV和hPVRalpha组成的囊泡以及具有降低的结合Tctex-1的能力的突变体hPVRalpha（hPVRMalpha）与Tctex-1共定位。然而，含有PV、右旋糖酐和hPVRalpha的囊泡仅具有逆行运动，而含有PV、右旋糖酐和hPVRMalpha的囊泡具有顺行或逆行运动。局部应用抗微管剂长春碱到坐骨神经减少了病毒从小腿运输到脊髓的数量。这些结果表明，直接有效的细胞质结构域和Tctex-1之间的相互作用是必不可少的有效逆行运输的PV-含有囊泡沿着微管在体内。
Poliovirus (PV), when injected intramuscularly into the calf, is incorporated into the sciatic nerve and causes an initial paralysis of the inoculated limb in transgenic mice carrying the human PV receptor (hPVR/CD155) gene. Here, we demonstrated by using an immunoelectron microscope that PV particles exist on vesicle structures in nerve terminals of neuromuscular junctions. We also demonstrated in glutathione S-transferase pull-down experiments that the dynein light chain, Tctex-1, interacts directly with the cytoplasmic domain of hPVR. In the axons of differentiated rat PC12 cells transfected with expression vectors for hPVRs, vesicles composed of PV and hPVRalpha, as well as a mutant hPVRalpha (hPVRMalpha) that had a reduced ability to bind Tctex-1, co-localized with Tctex-1. However, vesicles containing PV, dextran, and hPVRalpha had only retrograde motion, while those containing PV, dextran, and hPVRMalpha had anterograde or retrograde motion. Topical application of the antimicrotubule agent vinblastine to the sciatic nerve reduced the amount of virus transported from the calf to the spinal cord. These results suggest that direct efficient interaction between the cytoplasmic domain and Tctex-1 is essential for the efficient retrograde transport of PV-containing vesicles along microtubules in vivo.