Yamada H, Daikoku T, Yamashita Y, Jiang Y M, Tsurumi T, Nishiyama Y
Laboratory of Virology, Research Institute for Disease Mechanism and Control, Nagoya University School of Medicine, Japan.
J Gen Virol. 1997 Nov;78 ( Pt 11):2923-31. doi: 10.1099/0022-1317-78-11-2923.
We have identified the herpes simplex virus type 1 (HSV-1) US10 gene product using rabbit polyclonal antisera raised against a recombinant 6xHis-US10 fusion protein expressed in Escherichia coli. The antiserum reacted specifically with 34 and 36 kDa proteins in HSV-1 KOS-infected cells as shown by Western blotting and immunoprecipitation experiments. The 36 kDa protein was immunoprecipitated with the US10 antiserum from 32P-labelled lysates of Vero cells infected with HSV-1 KOS, demonstrating that the US10 protein was phosphorylated. Indirect immunofluorescence studies localized the US10 protein mainly to nuclei as large discrete particles at later times post-infection (p.i.), and nuclear fractionation studies revealed that the protein was tightly associated with the nuclear matrix. Moreover, analysis of isolated intracellular capsids showed that both phosphorylated and unphosphorylated forms of the US10 product were also associated with the capsid/tegument. These results indicate that the US10 gene of HSV-1 encodes a capsid/tegument-associated phosphoprotein which copurifies with the nuclear matrix.
我们使用针对在大肠杆菌中表达的重组6xHis-US10融合蛋白产生的兔多克隆抗血清，鉴定了单纯疱疹病毒1型（HSV-1）的US10基因产物。如蛋白质印迹和免疫沉淀实验所示，该抗血清与HSV-1 KOS感染细胞中的34 kDa和36 kDa蛋白发生特异性反应。用US10抗血清从感染HSV-1 KOS的Vero细胞的32P标记裂解物中免疫沉淀出36 kDa蛋白，表明US10蛋白被磷酸化。间接免疫荧光研究表明，在感染后较晚时间（p.i.），US10蛋白主要定位于细胞核，呈大的离散颗粒，细胞核分级分离研究表明该蛋白与核基质紧密相关。此外，对分离的细胞内衣壳的分析表明，US10产物的磷酸化和未磷酸化形式也与衣壳/包膜相关。这些结果表明，HSV-1的US10基因编码一种与衣壳/包膜相关的磷蛋白，该蛋白与核基质共纯化。
