Dong Chunsheng, Li Dan, Long Gang, Deng Fei, Wang Hualin, Hu Zhihong
State Key Laboratory of Virology, Key Laboratory of Molecular Virology and Joint-lab of Invertebrate Virology, Wuhan Institute of Virology, Chinese Academy of Sciences, Wuhan 430071, PR China.
Virology. 2005 Jul 20;338(1):112-20. doi: 10.1016/j.virol.2005.05.003.
Baculovirus encoded P10 form fibrillar structures in infected cells. We have tried to identify the functional domains for the P10 filament formation by green fluorescence protein (GFP) tag. The p10 gene of Helicoverpa armigera single nucleocapsid nucleopolyhedrovirus (HearNPV) was the subject of these studies. Different lengths of HearNPV p10 gene were constructed with GFP fused in frame to the C-terminus. The constructs were transfected into insect and mammalian cells and observed by confocal microscopy. The results indicated that the first N-terminal 66 amino acids, which form the complete coiled-coil domain, were necessary for the aggregation and formation of filament structures of HearNPV P10. The proline-rich region and the C-terminal positively charged amino acids were not necessary for the formation of fibrillar structure but had some impact on the shape of the fibrillar structures. No other baculoviral proteins were needed for the formation of P10 filament structures in transfected cells.
杆状病毒在受感染细胞中编码形成纤维状结构的P10。我们试图通过绿色荧光蛋白（GFP）标签鉴定P10丝状形成的功能域。棉铃虫单核衣壳核多角体病毒（HearNPV）的p10基因是这些研究的对象。构建了不同长度的HearNPV p10基因，并将GFP与C端框内融合。将构建体转染到昆虫和哺乳动物细胞中，并用共聚焦显微镜观察。结果表明，形成完整卷曲螺旋结构域的N端前66个氨基酸对于HearNPV P10丝状结构的聚集和形成是必需的。富含脯氨酸的区域和C端带正电荷的氨基酸对于纤维状结构的形成不是必需的，但对纤维状结构的形状有一定影响。在转染细胞中形成P10丝状结构不需要其他杆状病毒蛋白。
