2 OF THE 3 INFLUENZA VIRAL POLYMERASE PROTEINS EXPRESSED BY USING BACULOVIRUS VECTORS FORM A COMPLEX IN INSECT CELLS
2 OF THE 3 INFLUENZA VIRAL POLYMERASE PROTEINS EXPRESSED BY USING BACULOVIRUS VECTORS FORM A COMPLEX IN INSECT CELLS
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DOI：
10.1128/jvi.61.2.361-365.1987
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发表时间：
1987-02-01
影响因子：
5.4
通讯作者：
KRUG, RM
中科院分区：
文献类型：
--
作者：
STANGELO, C;SMITH, GE;KRUG, RM
将流感病毒聚合酶（P）基因PB 1、PB 2和PA中的每一个插入杆状病毒载体中，在多角体蛋白启动子的控制下。在昆虫（草地贪夜蛾）细胞感染的每一个杆状病毒重组含有P基因插入，大量的编码P蛋白的合成。感染细胞中总蛋白的凝胶电泳分析揭示了对应于编码的P蛋白的新蛋白条带的存在，该蛋白条带足够丰富，可以用考马斯蓝染色。在同时用PB 1和PB 2杆状病毒重组体感染的细胞中，形成了PB 1-PB 2复合物，该复合物用对PB 1或PB 2具有特异性的抗血清免疫沉淀。在同时感染所有三种P杆状病毒重组体的细胞中，形成了缺乏PA蛋白的PB 1-PB 2复合物。该PB 1-PB 2复合物的形成部分模拟了流感病毒感染细胞中发生的事件，其中所有三种P蛋白彼此形成复合物（B. M.德特延角圣安杰洛湾G. Katze和R. M. Krug，J. Virol. 61：16-22，1987）。这些结果表明，PB 1和PB 2形成复合物的能力是这两种蛋白质的内在特性，不需要其他流感病毒基因产物的参与。PA蛋白的免疫沉淀的P蛋白复合物在昆虫细胞感染的三个P杆状病毒重组体的情况下的可能原因进行了讨论。
Each of the influenza virus polymerase (P) genes PB1, PB2, and PA was inserted into a baculovirus vector under the control of the polyhedrin promoter. In insect (Spodoptera frugiperda) cells infected by each baculovirus recombinant containing a P gene insert, a large amount of the encoded P protein was synthesized. Gel electrophoretic analysis of the total proteins in infected cells revealed the presence of a new protein band corresponding to the encoded P protein that was abundant enough to be stained with Coomassie blue. In cells infected simultaneously with both the PB1 and PB2 baculovirus recombinants, a PB1-PB2 complex was formed that was immunoprecipitated with an antiserum specific for either PB1 or PB2. In cells infected simultaneously with all three P baculovirus recombinants, a PB1-PB2 complex lacking the PA protein was formed. Formation of this PB1-PB2 complex partially mimics events that occur in influenza virus-infected cells, where all three P proteins form a complex with each other (B. M. Detjen, C. St. Angelo, M. G. Katze, and R. M. Krug, J. Virol. 61:16-22, 1987). These results indicate that the ability of PB1 and PB2 to form a complex is an intrinsic property of these two proteins that does not require the participation of other influenza viral gene products. Possible reasons for the absence of the PA protein from the immunoprecipitable P protein complex in insect cells infected by the three P baculovirus recombinants are discussed.