Yi C K, Bansal O B, Hong M L, Chatterjee S, Roy P
Laboratory of Molecular Biophysics, University of Oxford, United Kingdom.
J Virol. 1996 Jul;70(7):4778-82. doi: 10.1128/JVI.70.7.4778-4782.1996.
Genome segment 9 of bluetongue virus serotype 10 encodes the minor protein VP6. The protein is abundant with basic residues particularly in two regions of the carboxy half of the molecule. A series of amino- and carboxy-terminal deletion mutants was expressed in mammalian cells by using a vaccinia virus T7 polymerase-driven transient expression system, and the intracellular fate of the products was monitored by both immunofluorescence staining and cell fractionation techniques. Data obtained indicated clearly that VP6 has nuclear transportation signals which may be correlated with positively charged domains of the molecule. In the intact molecule, though, these signals are masked and the protein is retained in the cytoplasm. The biochemical and immunofluorescence data obtained indicate that sequences in the region of residues 33 to 80 of the 328-amino acid protein are required for the retention of VP6 within the cell cytoplasm while amino acids 303 to 308 in the carboxy-terminal half of the molecule appear to possess nuclear localization capabilities.
蓝舌病毒血清型10的基因组片段9编码次要蛋白VP6。该蛋白富含碱性残基，尤其是在分子羧基端的两个区域。通过使用痘苗病毒T7聚合酶驱动的瞬时表达系统在哺乳动物细胞中表达了一系列氨基端和羧基端缺失突变体，并通过免疫荧光染色和细胞分级分离技术监测了产物在细胞内的命运。所获得的数据清楚地表明，VP6具有核运输信号，这些信号可能与分子的带正电荷结构域相关。然而，在完整分子中，这些信号被掩盖，蛋白质保留在细胞质中。所获得的生化和免疫荧光数据表明，328个氨基酸的蛋白质中33至80位残基区域的序列是VP6保留在细胞质中所必需的，而分子羧基端后半部分的303至308位氨基酸似乎具有核定位能力。
