Zhang S C, Wege C, Jeske H
Department of Molecular Biology and Plant Virology, Biological Institute, University of Stuttgart, Pfaffenwaldring 57, D-70550 Stuttgart, Germany.
Virology. 2001 Nov 25;290(2):249-60. doi: 10.1006/viro.2001.1185.
Two movement proteins (BV1 and BC1) facilitate the intra- and intercellular transport of begomoviruses in plants. In contrast to other geminiviruses the movement protein BC1 of Abutilon mosaic virus (AbMV) remained in the supernatant after centrifuging plant extracts at 20,000 g. To test whether this unusual behavior results from a distinct intracellular distribution of the protein, the BC1 gene has been fused to the gene of green fluorescent protein (GFP). The resulting plasmids were delivered into nonhost plants (Allium cepa) as well as into mature and immature cells of host plants (Nicotiana tabacum, N. benthamiana) by biolistic bombardment for transient expression in planta. BC1 directed GFP to two different cellular sites. In the majority of nonhost cells as well as in mature cells of host leaves, BC1 was mainly localized in small punctate flecks at the cell periphery or, to a lesser extent, around the nucleus. In sink leaves of host plants, GFP:BC1 additionally developed disc-like structures in the cell periphery. Cobombardment of GFP:BC1 with its cognate infectious DNA A and B did not change their subcellular distribution patterns in source leaves but led to the formation of peculiar needle-like structures in sink leaves. The nuclear shuttle protein (BV1) of AbMV accumulated mainly inside the nuclei as shown by immunohistochemical staining and GFP tagging. In sink cells of host plants it was mobilized to the plasma membrane and to the nucleus of the neighboring cell by coexpressed BC1, GFP:BC1, BC1:GFP, or after cobombardment with the cognate viral DNA. Only under these conditions were GFP:BC1 and BC1:GFP also found in the recipient cell.
两种运动蛋白(BV1和BC1)促进双生病毒在植物细胞内和细胞间的运输。与其他双生病毒不同,苘麻花叶病毒(AbMV)的运动蛋白BC1在以20,000 g离心植物提取物后仍留在上清液中。为了测试这种异常行为是否源于该蛋白独特的细胞内分布,已将BC1基因与绿色荧光蛋白(GFP)基因融合。通过生物弹轰击将所得质粒导入非寄主植物(洋葱)以及寄主植物(烟草、本氏烟草)的成熟和未成熟细胞中,以便在植物中瞬时表达。BC1将GFP导向两个不同的细胞位点。在大多数非寄主细胞以及寄主叶片的成熟细胞中,BC1主要定位于细胞周边的小斑点状区域,或在较小程度上定位于细胞核周围。在寄主植物的库叶中,GFP:BC1在细胞周边还形成了盘状结构。将GFP:BC1与其同源感染性DNA A和B共轰击,在源叶中并未改变它们的亚细胞分布模式,但在库叶中导致形成奇特的针状结构。AbMV的核穿梭蛋白(BV1)主要在细胞核内积累,免疫组织化学染色和GFP标记显示了这一点。在寄主植物的库细胞中,通过共表达BC1、GFP:BC1、BC1:GFP或与同源病毒DNA共轰击后,它被转运到质膜和相邻细胞细胞核中。只有在这些条件下,GFP:BC1和BC1:GFP也能在受体细胞中被发现。