Chattopadhyay D, Banerjee A K
Roche Institute of Molecular Biology, Roche Research Center, Nutley, NJ 07110.
Proc Natl Acad Sci U S A. 1987 Dec;84(24):8932-6. doi: 10.1073/pnas.84.24.8932.
The structural phosphoprotein NS of vesicular stomatitis virus, in association with the virion-associated RNA polymerase L protein, transcribes the genome ribonucleoprotein template in vitro. It contains an acidic N-terminal domain and two distinct domains at the C-terminal end that are involved in binding to the polymerase protein and the template RNA enwrapped with the nucleocapsid protein. In the present study, the portions of the NS gene that encode the N- and C-terminal domains of the protein were cloned in pGEM vectors and expressed by in vitro transcription and translation. It was shown that two polypeptides obtained by translation of the encoded mRNAs support RNA synthesis in vitro in a reconstitution reaction when they are added together in trans. Moreover, the N-terminal domain can be functionally substituted by structurally similar polypeptides.
水泡性口炎病毒的结构磷蛋白NS与病毒体相关的RNA聚合酶L蛋白一起，在体外转录基因组核糖核蛋白模板。它包含一个酸性N端结构域和C端的两个不同结构域，这两个结构域参与与聚合酶蛋白以及被核衣壳蛋白包裹的模板RNA的结合。在本研究中，编码该蛋白N端和C端结构域的NS基因部分被克隆到pGEM载体中，并通过体外转录和翻译进行表达。结果表明，由编码的mRNA翻译得到的两种多肽在体外重构反应中一起反式添加时支持RNA合成。此外，N端结构域可以被结构相似的多肽进行功能替代。
