Complex formation between influenza virus polymerase proteins expressed in Xenopus oocytes.
Complex formation between influenza virus polymerase proteins expressed in Xenopus oocytes.
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DOI：
10.1016/0042-6822(89)90523-0
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发表时间：
1989-07
期刊：
影响因子：
3.7
通讯作者：
Inglis SC
中科院分区：
文献类型：
--
作者：
Digard P;Blok VC;Inglis SC
所有三种流感病毒聚合酶 (P) 蛋白均通过显微注射的体外转录 mRNA 类似物在非洲爪蟾卵母细胞中表达，每个卵母细胞的产量高达 100 ng。为了检查爪蟾表达的 P 蛋白的功能状态，测试了多肽彼此形成稳定复合物的能力。正如在病毒感染的细胞中所见，所有三种 P 蛋白均结合成免疫沉淀复合物，表明该系统对于重建活性流感 RNA 聚合酶具有相当大的前景。对 P 蛋白配对组合缔合能力的检查表明，PB1 含有 PB2 和 PA 的独立结合位点，因此可能形成复合物的主链。游离和复合 P 蛋白的沉降分析表明 PB1 和 PB2 并不作为游离单体存在，并且类似地，所有三种 P 蛋白的复合物并不简单地由每种蛋白的一个拷贝组成。所有三种 P 蛋白复合物的异分散沉降速率似乎并不是由它们与 RNA 的结合引起的，这表明掺入了额外的多肽聚合酶复合物。
All three influenza virus polymerase (P) proteins were expressed in Xenopus oocytes from microinjected in vitro transcribed mRNA analogs, with yields of up to 100 ng per oocyte. To examine the functional state of the Xenopus-expressed P proteins, the polypeptides were tested for their ability to form stable complexes with each other. As seen in virus-infected cells, all three P proteins associated into an immunoprecipitable complex, suggesting that the system has considerable promise for the reconstruction of an active influenza RNA polymerase. Examination of the ability of paired combinations of the P proteins to associate indicated that PB1 contained independent binding sites for PB2 and PA, and so probably formed the backbone of the complex. Sedimentation analysis of free and complexed P proteins indicated that PB1 and PB2 did not exist as free monomers, and that similarly, complexes of all three P proteins did not simply consist of one copy of each protein. The heterodisperse sedimentation rate seen for complexes of all three P proteins did not appear to result from their binding to RNA, suggesting the incorporation of additional polypeptides polymerase complex.
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DOI：
10.1073/pnas.83.8.2709
发表时间：
1986-04-01
影响因子：
11.1
作者：
KRYSTAL, M;RUAN, L;PALESE, P
通讯作者：
PALESE, P
DOI：
10.1073/pnas.76.9.4350
发表时间：
1979-01-01
影响因子：
11.1
作者：
TOWBIN, H;STAEHELIN, T;GORDON, J
通讯作者：
GORDON, J
影响因子：
14.9
作者：
KRIEG, PA;MELTON, DA
通讯作者：
MELTON, DA
影响因子：
5.4
作者：
STANGELO, C;SMITH, GE;KRUG, RM
通讯作者：
KRUG, RM
影响因子：
11.4
作者：
Brierley, I;Boursnell, M E;Binns, M M;Bilimoria, B;Blok, V C;Brown, T D;Inglis, S C
通讯作者：
Inglis, S C