Pascal E, Sanderfoot A A, Ward B M, Medville R, Turgeon R, Lazarowitz S G
Department of Microbiology, University of Illinois at Urbana-Champaign 61801.
Plant Cell. 1994 Jul;6(7):995-1006. doi: 10.1105/tpc.6.7.995.
Plant viruses encode movement proteins that are essential for infection of the host but are not required for viral replication or encapsidation. Squash leaf curl virus (SqLCV), a bipartite geminivirus with a single-stranded DNA genome, encodes two movement proteins, BR1 and BL1, that have been implicated in separate functions in viral movement. To further elucidate these functions, we have investigated the nucleic acid binding properties and cellular localization of BR1 and BL1. In this study, we showed that BR1 binds strongly to single-stranded nucleic acids, with a higher affinity for single-stranded DNA than RNA, and is localized to the nucleus of SqLCV-infected plant cells. In contrast, BL1 binds only weakly to single-stranded nucleic acids and not at all to double-stranded DNA. The nuclear localization of BR1 and the previously demonstrated plasma membrane localization of BL1 were also observed when these proteins were expressed from baculovirus vectors in Spodoptera frugiperda insect cells. The biochemical properties and cellular locations of BR1 and BL1 suggest a model for SqLCV movement whereby BR1 is involved in the shuttling of the genome in and/or out of the nucleus and BL1 acts at the plasma membrane/cell wall to facilitate viral movement across cell boundaries.
植物病毒编码运动蛋白,这些蛋白对于感染宿主至关重要,但病毒复制或衣壳化过程并不需要它们。南瓜曲叶病毒(SqLCV)是一种具有单链DNA基因组的双分体双生病毒,它编码两种运动蛋白BR1和BL1,这两种蛋白在病毒运动中具有不同的功能。为了进一步阐明这些功能,我们研究了BR1和BL1的核酸结合特性及细胞定位。在本研究中,我们发现BR1与单链核酸紧密结合,对单链DNA的亲和力高于RNA,并且定位于被SqLCV感染的植物细胞的细胞核中。相比之下,BL1仅与单链核酸微弱结合,与双链DNA完全不结合。当这些蛋白在草地贪夜蛾昆虫细胞中由杆状病毒载体表达时,也观察到了BR1的细胞核定位以及之前证明的BL1的质膜定位。BR1和BL1的生化特性及细胞定位提示了一种SqLCV运动模型,即BR1参与基因组进出细胞核的穿梭过程,而BL1在质膜/细胞壁发挥作用,促进病毒跨细胞边界移动。