Nakagawa Y, Oda K, Nakada S
Department of Biological Science and Technology, Science University of Tokyo, Chiba, Japan.
J Virol. 1996 Sep;70(9):6390-4. doi: 10.1128/JVI.70.9.6390-6394.1996.
We indicated that the PB1 and PA subunits of RNA polymerase and nucleoprotein (NP) can support replication of the influenza virus genome as well as transcription to yield uncapped poly(A)(+)-RNA (Y. Nakagawa, N. Kimura, T. Toyoda, K. Mizumoto, A. Ishihama, K. Oda, and S. Nakada, J. Virol. 69:728-733, 1995). To analyze the functions of the PB1 and PA subunits in replication and transcription, YP1N clones in which the PB1 and NP genes can be expressed in response to dexamethasone were established. cRNA was transcribed from model viral RNA (vRNA), but vRNA synthesis from model cRNA was not detected in YP1N clones. Furthermore, poly(A)(+)-RNA directed from model vRNA was synthesized in YP1N clones. These results indicated that PB1 and NP can support the syntheses of cRNA and poly(A)(+)-RNA and that the PA subunit, in addition to that of PB1 and to NP, is required for vRNA synthesis. In summary, the PB1 subunit is involved in the catalytic activities of nucleotide elongation, and the PA subunit may act as an allosteric modulator and cause a conformational change from a cRNA-to a vRNA-synthesizing form of the PB1 subunit.
我们指出,RNA聚合酶的PB1和PA亚基以及核蛋白(NP)能够支持流感病毒基因组的复制以及转录,从而产生无帽的多聚腺苷酸(+)-RNA(Y. 中川、N. 木村、T. 丰田、K. 水本、A. 石滨、K. 小田和S. 中田,《病毒学杂志》69:728 - 733,1995年)。为了分析PB1和PA亚基在复制和转录中的功能,构建了YP1N克隆,其中PB1和NP基因可响应地塞米松进行表达。从模型病毒RNA(vRNA)转录生成了cRNA,但在YP1N克隆中未检测到从模型cRNA合成vRNA。此外,在YP1N克隆中合成了由模型vRNA指导的多聚腺苷酸(+)-RNA。这些结果表明,PB1和NP能够支持cRNA和多聚腺苷酸(+)-RNA的合成,并且除了PB1和NP外,PA亚基对于vRNA合成也是必需的。总之,PB1亚基参与核苷酸延伸的催化活性,而PA亚基可能作为变构调节剂,使PB1亚基从合成cRNA的构象转变为合成vRNA的构象。