Rojas M R, Zerbini F M, Allison R F, Gilbertson R L, Lucas W J
Division of Biological Sciences, University of California, Davis, California 95616, USA.
Virology. 1997 Oct 27;237(2):283-95. doi: 10.1006/viro.1997.8777.
The role of bean common mosaic necrosis potyvirus (BCMNV) and lettuce mosaic potyvirus (LMV) proteins was investigated in terms of their capacity to function as viral movement proteins (MPs). Using Escherichia coli-expressed proteins and microinjection techniques, direct evidence was obtained that both the potyviral capsid protein (CP) and helper component- proteinase (HC-Pro) function in this capacity, in that both proteins (a) trafficked from cell to cell, (b) induced an increase in plasmodesmal size exclusion limit, and (c) facilitated cell-to-cell movement of viral RNA. CP and HC-Pro mutants were also produced and used in microinjection experiments. Mutations in the core region of the CP either impaired (single and double amino acid substitution mutants) or abolished (triple amino acid substitution mutant) cell-to-cell movement, as did C-terminal deletion mutants in HC-Pro. The BCMNV P1, CI, NIa, and NIb proteins did not exhibit viral MP properties, but NIa and NIb proteins were found to accumulate within the nuclei of injected cells. These results further establish the multifunctional nature of the potyvirus CP and HC-Pro.
研究了菜豆普通花叶坏死病毒(BCMNV)和莴苣花叶病毒(LMV)蛋白作为病毒运动蛋白(MPs)发挥功能的能力。利用大肠杆菌表达的蛋白和显微注射技术,获得了直接证据,证明这两种马铃薯Y病毒属病毒的衣壳蛋白(CP)和辅助成分蛋白酶(HC-Pro)均具有此功能,即这两种蛋白(a)能在细胞间运输,(b)能诱导胞间连丝大小排阻极限增加,(c)能促进病毒RNA的细胞间移动。还构建了CP和HC-Pro突变体并用于显微注射实验。CP核心区域的突变要么损害(单氨基酸和双氨基酸取代突变体)要么消除(三氨基酸取代突变体)细胞间移动,HC-Pro的C末端缺失突变体也有同样情况。BCMNV的P1、CI、NIa和NIb蛋白未表现出病毒MP特性,但发现NIa和NIb蛋白在注射细胞的细胞核内积累。这些结果进一步证实了马铃薯Y病毒属病毒CP和HC-Pro的多功能性质。