Fedorkin O N, Merits A, Lucchesi J, Solovyev A G, Saarma M, Morozov S Y, Mäkinen K
Institute of Biotechnology, Program for Plant Molecular Biology, Viikki Biocentre, University of Helsinki, (Viikinkaari 9), Helsinki, FIN-00014, Finland.
Virology. 2000 Apr 25;270(1):31-42. doi: 10.1006/viro.2000.0246.
The cell-to-cell movement of the GUS-tagged potato virus X (PVX) coat protein (CP) movement-deficient mutant was restored by potyviral CPs of potato virus A (PVA) and potato virus Y (PVY) in Nicotiana benthamiana leaves in transient cobombardment experiments. Viral cell-to-cell movement of PVX CP mutant was complemented in Nicotiana tabacum cv. SR1 transgenic plants expressing PVY CP: PVX RNA and polymerase were detected in the PVX CP mutant-inoculated leaves of transgenic plants. These findings demonstrated the ability of the PVX CP-deficient mutant to move from cell to cell but not long distances in the transgenic plants and suggest that CPs of potex- and potyviruses display complementary activities in the movement process. Potyviral CP alone is not able to carry out these activities, since the mutated PVX CP is indispensable for restored movement. No trans-encapsidation between potyviral CP and PVX RNA was observed. Therefore, potyviral CP facilitates the PVX CP mutant movement by the mechanism that cannot be explained by coat protein substitution. Our data also suggest that CP functioning in cell-to-cell movement is not restricted to a simple passive role in forming virions.
在瞬时共轰击实验中,马铃薯A病毒(PVA)和马铃薯Y病毒(PVY)的马铃薯Y病毒属病毒外壳蛋白(CP)可恢复带有GUS标签的马铃薯X病毒(PVX)外壳蛋白运动缺陷型突变体在本氏烟草叶片中的细胞间移动。PVX CP突变体的病毒细胞间移动在表达PVY CP的烟草品种SR1转基因植物中得到互补:在转基因植物接种PVX CP突变体的叶片中检测到PVX RNA和聚合酶。这些发现证明了PVX CP缺陷型突变体在转基因植物中具有细胞间移动能力,但不能进行长距离移动,这表明马铃薯X病毒属病毒和马铃薯Y病毒属病毒的外壳蛋白在移动过程中表现出互补活性。单独的马铃薯Y病毒属病毒CP无法进行这些活动,因为突变的PVX CP对于恢复移动是不可或缺的。未观察到马铃薯Y病毒属病毒CP与PVX RNA之间的转衣壳现象。因此,马铃薯Y病毒属病毒CP通过无法用外壳蛋白替代来解释的机制促进PVX CP突变体的移动。我们的数据还表明,在细胞间移动中起作用的CP不限于在形成病毒粒子中扮演简单的被动角色。