Three-dimensional model for the isolated recombinant influenza virus polymerase heterotrimer.
Three-dimensional model for the isolated recombinant influenza virus polymerase heterotrimer.
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DOI：
10.1093/nar/gkm336
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发表时间：
2007
影响因子：
14.9
通讯作者：
Llorca O
中科院分区：
文献类型：
--
作者：
Torreira E;Schoehn G;Fernández Y;Jorba N;Ruigrok RW;Cusack S;Ortín J;Llorca O
甲型流感病毒的基因组被组织成八个核糖核蛋白复合物（RNP），每个RNP含有一个RNA聚合酶复合物。这种RNA聚合酶也被发现与RNP无关，并且可能参与感染周期中的不同功能。我们通过共转染PB1、PB2和PA基因在哺乳动物细胞中表达了病毒RNA聚合酶复合物，并通过TAP标签程序纯化了异源三聚体。通过电子显微镜和单粒子图像处理确定其三维结构。所获得的模型类似于先前报道的与病毒RNP相关的聚合酶复合物的结构，但似乎处于更开放的构象。详细的模型比较表明，复杂的特定区域显示重要的构象变化相比，RNP相关的聚合酶的结构，特别是在已知的区域与相邻的NP单体的RNP相互作用。此外，PB2亚基似乎由于聚合酶与RNP的缔合而经历了实质性的置换。提出的结构模型表明，在溶液中的聚合酶的核心构象存在，但与其他合作伙伴，如蛋白质或RNA的相互作用，将触发不同的构象变化，激活新的功能特性。
The genome of influenza A virus is organized into eight ribonucleoprotein complexes (RNPs), each containing one RNA polymerase complex. This RNA polymerase has also been found non-associated to RNPs and is possibly involved in distinct functions in the infection cycle. We have expressed the virus RNA polymerase complex by co-tranfection of the PB1, PB2 and PA genes in mammalian cells and the heterotrimer was purified by the TAP tag procedure. Its 3D structure was determined by electron microscopy and single-particle image processing. The model obtained resembles the structure previously reported for the polymerase complex associated to viral RNPs but appears to be in a more open conformation. Detailed model comparison indicated that specific areas of the complex show important conformational changes as compared to the structure for the RNP-associated polymerase, particularly in regions known to interact with the adjacent NP monomers in the RNP. Also, the PB2 subunit seems to undergo a substantial displacement as a result of the association of the polymerase to RNPs. The structural model presented suggests that a core conformation of the polymerase in solution exists but the interaction with other partners, such as proteins or RNA, will trigger distinct conformational changes to activate new functional properties.
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